46 results on '"Susan Idicula-Thomas"'
Search Results
2. Expanding the therapeutic options for Candida infections using novel inhibitors of secreted aspartyl proteases
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Shuvechha Chakraborty, Kshitija Rahate, Chandan Kumar, and Susan Idicula‐Thomas
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Drug Discovery - Abstract
For widening the therapeutic options for Candida management, the druggability of Candida proteome was systematically investigated using an innovative pipeline of high-throughput data mining algorithms, followed by in vitro validation of the observations. Through this exercise, HIV-1 protease was found to share structural similarity with secreted aspartyl protease-3 (SAP3), a virulence protein of Candida. Using the molecular fingerprint of HIV-1 protease inhibitor GRL-09510, we performed virtual screening of peptidomimetic library followed by high-precision docking and MD simulations for discovery of SAP inhibitors. Wet-lab validation of the four shortlisted peptidomimetics revealed that two molecules, when used in combination with fluconazole, could significantly reduce the dosage of fluconazole required for 50% inhibition of Candida albicans. The SAP inhibitory activity of these peptidomimetics was confirmed through SAP assays and found to be on par with pepstatin A, a known peptidomimetic inhibitor of aspartyl proteases.
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- 2022
3. CAMPR4: a database of natural and synthetic antimicrobial peptides
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Ulka Gawde, Shuvechha Chakraborty, Faiza Hanif Waghu, Ram Shankar Barai, Ashlesha Khanderkar, Rishikesh Indraguru, Tanmay Shirsat, and Susan Idicula-Thomas
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Genetics - Abstract
There has been an exponential increase in the design of synthetic antimicrobial peptides (AMPs) for its use as novel antibiotics. Synthetic AMPs are substantially enriched in residues with physicochemical properties known to be critical for antimicrobial activity; such as positive charge, hydrophobicity, and higher alpha helical propensity. The current prediction algorithms for AMPs have been developed using AMP sequences from natural sources and hence do not perform well for synthetic peptides. In this version of CAMP database, along with updating sequence information of AMPs, we have created separate prediction algorithms for natural and synthetic AMPs. CAMPR4 holds 24243 AMP sequences, 933 structures, 2143 patents and 263 AMP family signatures. In addition to the data on sequences, source organisms, target organisms, minimum inhibitory and hemolytic concentrations, CAMPR4 provides information on N and C terminal modifications and presence of unusual amino acids, as applicable. The database is integrated with tools for AMP prediction and rational design (natural and synthetic AMPs), sequence (BLAST and clustal omega), structure (VAST) and family analysis (PRATT, ScanProsite, CAMPSign). The data along with the algorithms of CAMPR4 will aid to enhance AMP research. CAMPR4 is accessible at http://camp.bicnirrh.res.in/.
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- 2022
4. Pathway Analysis of Genome Wide Association Studies (GWAS) Data Associated with Male Infertility
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Rupashree Salvi, Ulka Gawde, Susan Idicula-Thomas, and Barnali Biswas
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Geology ,Ocean Engineering ,Water Science and Technology - Abstract
Background: Infertility is a common condition affecting approximately 10–20% of the reproductive age population. Idiopathic infertility cases are thought to have a genetic basis, but the underlying causes are largely unknown. However, the genetic basis underlying male infertility in humans is only partially understood. The Purpose of the study is to understand the current state of research on the genetics of male infertility and its association with significant biological mechanisms. Results: We performed an Identify Candidate Causal SNPs and Pathway (ICSN Pathway) analysis using a genome-wide association study (GWAS) dataset, and NCBI-PubMed search which included 632 SNPs in GWAS and 451 SNPs from the PubMed server, respectively. The ICSN Pathway analysis produced three hypothetical biological mechanisms associated with male infertility: (1) rs8084 and rs7192→HLA-DRA→inflammatory pathways and cell adhesion; rs7550231 and rs2234167→TNFRSF14→TNF Receptor Superfamily Member 14→T lymphocyte proliferation and activation; rs1105879 and rs2070959→UGT1A6→UDP glucuronosyltransferase family 1 member A6→Metabolism of Xenobiotics, androgen, estrogen, retinol, and carbohydrates. Conclusions: We believe that our results may be helpful to study the genetic mechanisms of male infertility. Pathway-based methods have been applied to male infertility GWAS datasets to investigate the biological mechanisms and reported some novel male infertility risk pathways. This pathway analysis using GWAS dataset suggests that the biological process related to inflammation and metabolism might contribute to male infertility susceptibility. Our analysis suggests that genetic contribution to male infertility operates through multiple genes affecting common inflammatory diseases interacting in functional pathways.
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- 2022
5. A Recombinant Fragment of Human Surfactant Protein D Binds Spike Protein and Inhibits Infectivity and Replication of SARS-CoV-2 in Clinical Samples
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Uday Kishore, Ekta Gupta, Hrishikesh Pandit, Indra Kundu, Sheetalnath Rooge, Savneet Kaur, Reshu Agarwal, Sanjeev Gupta, Taruna Madan, Dinesh M. Tripathi, Susan Idicula-Thomas, Praveen M. Varghese, Rambhadur Subedi, and Barnali Biswas
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entry inhibition ,Male ,0301 basic medicine ,viruses ,Clinical Biochemistry ,Plasma protein binding ,Virus Replication ,spike protein ,medicine.disease_cause ,law.invention ,0302 clinical medicine ,law ,Chlorocebus aethiops ,Receptor ,skin and connective tissue diseases ,Pulmonary Surfactant-Associated Protein D ,Original Research ,Coronavirus ,Infectivity ,Chemistry ,virus diseases ,Recombinant Proteins ,Spike Glycoprotein, Coronavirus ,Recombinant DNA ,Female ,Protein Binding ,medicine.drug ,Adult ,Pulmonary and Respiratory Medicine ,surfactant protein D ,Protein subunit ,Virus ,03 medical and health sciences ,medicine ,Animals ,Humans ,Gene ,Vero Cells ,Molecular Biology ,entry inhibitor ,SARS-CoV-2 ,fungi ,COVID-19 ,Surfactant protein D ,Cell Biology ,Molecular biology ,Virology ,Entry inhibitor ,body regions ,030104 developmental biology ,030228 respiratory system ,Viral replication - Abstract
RationaleCOVID-19 is an acute infectious disease caused by the Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2). Human surfactant protein D (SP-D) is known to interact with spike protein of SARS-CoV, but its immune-surveillance against SARS-CoV-2 is not known.ObjectiveThis study aimed to examine the potential of a recombinant fragment of human SP-D (rfhSP-D) as an inhibitor of replication and infection of SARS-CoV-2.MethodsrfhSP-D interaction with spike protein of SARS-CoV-2 and hACE-2 receptor was predicted via docking analysis. The inhibition of interaction between spike protein and ACE-2 by rfhSP-D was confirmed using direct and indirect ELISA. The effect of rfhSP-D on replication and infectivity of SARS-CoV-2 from clinical samples was studied by measuring the expression of RdRp gene of the virus using qPCR.Measurements and Main ResultsIn-silico interaction studies indicated that three amino acid residues in the RBD of spike of SARS-CoV-2 were commonly involved in interacting with rfhSP-D and ACE-2. Studies using clinical samples of SARS-CoV-2 positive cases (asymptomatic, n=7 and symptomatic, n=8 and negative controls n=15) demonstrated that treatment with 5μM rfhSP-D inhibited viral replication by ~5.5 fold and was more efficient than Remdesivir (100 μM). Approximately, a 2-fold reduction in viral infectivity was also observed after treatment with 5μM rfhSP-D.ConclusionsThese results conclusively demonstrate that the calcium independent rfhSP-D mediated inhibition of binding between the receptor binding domain of the S1 subunit of the SARS-CoV-2 spike protein and human ACE-2, its host cell receptor, and a significant reduction in SARS-CoV-2 infection and replication in-vitro.
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- 2021
6. Exploring the druggable proteome of Candida species through comprehensive computational analysis
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Shuvechha Mukherjee, Susan Idicula-Thomas, Mehdi Askari, Indra Kundu, Ram Shankar Barai, and Kareenhalli V. Venkatesh
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0106 biological sciences ,Antifungal Agents ,Proteome ,In silico ,Systems biology ,Druggability ,Genomics ,Computational biology ,Secondary metabolite ,01 natural sciences ,Fungal Proteins ,Candida tropicalis ,03 medical and health sciences ,Candida albicans ,Drug Discovery ,Genetics ,medicine ,030304 developmental biology ,0303 health sciences ,biology ,Drug Repositioning ,Computational Biology ,Bridged Bicyclo Compounds, Heterocyclic ,biology.organism_classification ,Metabolic Flux Analysis ,Diterpenes ,Software ,Protein Binding ,010606 plant biology & botany ,medicine.drug - Abstract
Candida albicans and non-albicans Candida spp. are major cause of systemic mycoses. Antifungal drugs such as azoles and polyenes are not efficient to successfully eradicate Candida infection owing to their fungistatic nature or low bioavailability. Here, we have adopted a comprehensive computational workflow for identification, prioritization and validation of targets from proteomes of Candida albicans and Candida tropicalis. The protocol involves identification of essential drug-target candidates using subtractive genomics, protein-protein interaction network properties and systems biology based methods. The essentiality of the novel metabolic and non-metabolic targets was established by performing in silico gene knockouts, under aerobic as well as anaerobic conditions, and in vitro drug inhibition assays respectively. Deletion of twelve genes that are involved in amino acid, secondary metabolite, and carbon metabolism showed zero growth in metabolic model under simulated conditions. The algorithm, used in this study, can be downloaded from http://pbit.bicnirrh.res.in/offline.php and executed locally.
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- 2021
7. FSHR activation through small molecule modulators: Mechanistic insights from MD simulations
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Chandan Kumar and Susan Idicula-Thomas
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Health Informatics ,Computer Science Applications - Published
- 2023
8. Multimorbidity landscape of schizophrenia: Insights from meta-analysis of genome wide association studies
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Ulka Gawde, Priyanka Hegde, Karishma Desai, Ram Shankar Barai, Bipin Raj Shekhar, Dhanjit K. Das, and Susan Idicula-Thomas
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Psychiatry and Mental health ,Schizophrenia ,Humans ,Multimorbidity ,Genetic Predisposition to Disease ,Polymorphism, Single Nucleotide ,Biological Psychiatry ,Genome-Wide Association Study - Published
- 2022
9. Comparison of machine learning algorithms applied to symptoms to determine infectious causes of death in children: national survey of 18,000 verbal autopsies in the Million Death Study in India
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Susan Idicula-Thomas, Ulka Gawde, and Prabhat Jha
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Child mortality ,medicine.medical_specialty ,India ,Cause of death ,Machine learning ,computer.software_genre ,Million Death Study ,Communicable Diseases ,Measles ,Machine Learning ,Prediction model ,Epidemiology ,medicine ,Humans ,Verbal autopsy ,Fever of unknown origin ,Child ,Infectious disease ,business.industry ,Research ,Public Health, Environmental and Occupational Health ,Infant ,medicine.disease ,Child, Preschool ,Autopsy ,Artificial intelligence ,Public aspects of medicine ,RA1-1270 ,business ,Meningitis ,Algorithm ,computer ,Algorithms ,Encephalitis - Abstract
Background Machine learning (ML) algorithms have been successfully employed for prediction of outcomes in clinical research. In this study, we have explored the application of ML-based algorithms to predict cause of death (CoD) from verbal autopsy records available through the Million Death Study (MDS). Methods From MDS, 18826 unique childhood deaths at ages 1–59 months during the time period 2004–13 were selected for generating the prediction models of which over 70% of deaths were caused by six infectious diseases (pneumonia, diarrhoeal diseases, malaria, fever of unknown origin, meningitis/encephalitis, and measles). Six popular ML-based algorithms such as support vector machine, gradient boosting modeling, C5.0, artificial neural network, k-nearest neighbor, classification and regression tree were used for building the CoD prediction models. Results SVM algorithm was the best performer with a prediction accuracy of over 0.8. The highest accuracy was found for diarrhoeal diseases (accuracy = 0.97) and the lowest was for meningitis/encephalitis (accuracy = 0.80). The top signs/symptoms for classification of these CoDs were also extracted for each of the diseases. A combination of signs/symptoms presented by the deceased individual can effectively lead to the CoD diagnosis. Conclusions Overall, this study affirms that verbal autopsy tools are efficient in CoD diagnosis and that automated classification parameters captured through ML could be added to verbal autopsies to improve classification of causes of death.
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- 2021
10. Mapping of FSHR agonists and antagonists binding sites to identify potential peptidomimetic modulators
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Muthu Sankar Aathi, Chandan Kumar, Kaushiki S. Prabhudesai, Dhivya Shanmugarajan, and Susan Idicula-Thomas
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Binding Sites ,Databases, Factual ,Biophysics ,Cell Biology ,Molecular Dynamics Simulation ,Biochemistry ,Molecular Docking Simulation ,HEK293 Cells ,Allosteric Regulation ,Protein Domains ,Peptide Library ,Humans ,Receptors, FSH ,Peptidomimetics - Abstract
Owing to the critical role of follicle stimulating hormone receptor (FSHR) signaling in human reproduction, FSHR has been widely explored for development of fertility regulators. Using high-throughput screening approaches, several low molecular weight (LMW) compounds that can modulate FSHR activity have been identified. However, the information about the binding sites of these molecules on FSHR is not known. In the present study, we extracted the structural and functional information of 161 experimentally validated LMW FSHR modulators available in PubMed records. The potential FSHR binding sites for these modulators were identified through molecular docking experiments. The binding sites were further mapped to the agonist or antagonist activity reported for these molecules in literature. MD simulations were performed to evaluate the effect of ligand binding on conformational changes in the receptor, specifically the transmembrane domain. A peptidomimetic library was screened using these binding sites. Six peptidomimetics that interacted with the residues of transmembrane domain and extracellular loops were evaluated for binding activity using in vitro cAMP assay. Two of the six peptidomimetics exhibited positive allosteric modulatory activity and four peptidomimetics exhibited negative allosteric modulatory activity. All six peptidomimetics interacted with Asp521 of hFSHR(TMD). Several of the experimentally known LMW FSHR modulators also participated in H-bond interactions with Asp521, suggesting its important role in FSHR modulatory activity.
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- 2021
11. Collection of antimicrobial peptides database and its derivatives: Applications and beyond
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Faiza Hanif Waghu and Susan Idicula-Thomas
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Antifungal ,0303 health sciences ,Tools for Protein Science ,Structure analysis ,medicine.drug_class ,Computer science ,030302 biochemistry & molecular biology ,Antimicrobial peptides ,Computational Biology ,Molecular Sequence Annotation ,Computational biology ,Cloud Computing ,Biochemistry ,Markov Chains ,Anti-Bacterial Agents ,Machine Learning ,03 medical and health sciences ,Identification (information) ,medicine ,Amino Acid Sequence ,Databases, Protein ,Peptides ,Hidden Markov model ,Molecular Biology ,030304 developmental biology - Abstract
Collection of antimicrobial peptides (CAMP), CAMPSign, and ClassAMP are open‐access resources that have been developed to enhance research on antimicrobial peptides (AMPs). Comprehensive information on AMPs and machine learning‐based predictive models are made available for users through these resources. As of date, CAMP(R3) has 10,247 sequences, 757 structures, and 114 family‐specific signatures of AMPs along with associated tools for AMP sequence and structure analysis. CAMPSign uses family‐specific sequence conservation, in the form of patterns and hidden Markov models for identification of AMPs. ClassAMP can be used to classify AMPs as antibacterial, antifungal, or antiviral based on sequence information. Here we describe CAMP and its derivatives and illustrate, with a few examples, the contribution of these online resources to the advancement of our current understanding of AMPs.
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- 2019
12. Discovery of small molecule binders of human FSHR(TMD) with novel structural scaffolds by integrating structural bioinformatics and machine learning algorithms
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Sanchi Shah, Bhawana Sahu, Susan Idicula-Thomas, Kaushiki S. Prabhudesai, and Alessandro Contini
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Models, Molecular ,0301 basic medicine ,endocrine system ,Quantitative structure–activity relationship ,Allosteric regulation ,Quantitative Structure-Activity Relationship ,Computational biology ,Ligands ,01 natural sciences ,Cell Line ,Machine Learning ,Computers, Molecular ,03 medical and health sciences ,Structural bioinformatics ,0103 physical sciences ,Materials Chemistry ,Humans ,Physical and Theoretical Chemistry ,Spectroscopy ,Virtual screening ,Binding Sites ,010304 chemical physics ,Chemistry ,Computational Biology ,Computer Graphics and Computer-Aided Design ,Small molecule ,Transmembrane domain ,030104 developmental biology ,Docking (molecular) ,Drug Design ,Receptors, FSH ,Follicle-stimulating hormone receptor ,Hydrophobic and Hydrophilic Interactions ,Protein Binding - Abstract
Background The activation of follicle stimulating hormone receptor (FSHR) by FSH and the consequent downstream signaling activities are crucial for reproductive health. The role of FSHR in tumor progression as well as osteoporosis advancement has also been well established. Currently, steroid preparations of estrogen and progesterone are being used for managing fertility, in spite of the harmful side effects, as there has not been much success in identification of effective FSHR modulators. Structure-based drug design initiatives for identification of potent and specific FSHR modulators have been impeded due to the non-availability of the complete crystal structure of hFSHR complexed with FSH. Methods In this study, we have modeled the 3D structure of transmembrane domain (TMD) of hFSHR and identified molecules that demonstrate good binding affinity by virtual screening of drug-like library of compounds. The 3D structural and pharmacophoric features of the binders and non-binders obtained from virtual screening were further used to develop Support Vector Machine based classifier for TMD binding. Based on the observations from docking and SVM classification, a small molecule was identified for extensive MD simulations and in vitro assays for FSHR modulatory activity. Results The molecule selected based on docking score and SVM prediction was found to inhibit FSH-induced cAMP activity by 80% at 300 μM concentration. Conclusion The study proposes 1,3-diphenyl-1H-pyrazole-5-carboxylate as a promising scaffold for the design of new and potent FSHR allosteric inhibitors.
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- 2019
13. Central residues of FSHβ (89-97) peptide are not critical for FSHR binding: Implications for peptidomimetic design
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Karishma Desai, Susan Idicula-Thomas, Vikas Dighe, Kaushiki S. Prabhudesai, Deepak Modi, Alessandro Contini, and Sahil Raje
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Models, Molecular ,endocrine system ,Peptidomimetic ,medicine.drug_class ,Clinical Biochemistry ,Pharmaceutical Science ,Peptide ,01 natural sciences ,Biochemistry ,Downregulation and upregulation ,Drug Discovery ,medicine ,Animals ,Humans ,Molecular Biology ,Granulosa cell proliferation ,chemistry.chemical_classification ,Alanine ,Binding Sites ,010405 organic chemistry ,Organic Chemistry ,Ovary ,Antagonist ,0104 chemical sciences ,Cell biology ,Androgen receptor ,010404 medicinal & biomolecular chemistry ,chemistry ,Estrogen ,Drug Design ,Molecular Medicine ,Receptors, FSH ,Female ,Peptidomimetics ,Follicle Stimulating Hormone ,Peptides - Abstract
In our previous study, we had identified a 9-mer peptide (FSHβ (89–97)) derived from seat belt loop of human FSHβ and demonstrated its ability to function as FSHR antagonist in vivo. Structure analysis revealed that the four central residues 91STDC94 within this peptide may not be critical for receptor binding. In the present study, 91STDC94 residues were substituted with alanine to generate ΔFSHβ 89–97(91STDC94/AAAA) peptide. Analogous to the parent peptide, ΔFSHβ 89–97(91STDC94/AAAA) peptide inhibited binding of iodinated FSH to rat FSHR and reduced FSH-induced cAMP production. The peptide could impede granulosa cell proliferation leading to reduction in FSH-mediated ovarian weight gain in immature female rats. In these rats, peptide administration further downregulated androgen receptor and estrogen receptor-alpha expression and upregulated estrogen receptor-beta expression. The results indicate that substitution of 91STDC94 with alanine did not significantly alter FSHR antagonist activity of FSHβ (89–97) peptide implying that these residues are not critical for FSH-FSHR interaction and can be replaced with non-peptidic moieties for development of more potent peptidomimetics.
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- 2021
14. Enrichment analyses of diseases and pathways associated with precocious puberty using PrecocityDB
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Sameeksha Bhaye, Karishma Desai, Ram Shankar Barai, Khushal Pokar, Susan Idicula-Thomas, Mridula Sharma, and Indra Kundu
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0301 basic medicine ,Databases, Factual ,Science ,Puberty, Precocious ,030209 endocrinology & metabolism ,Single-nucleotide polymorphism ,Comorbidity ,Disease ,Biology ,Polymorphism, Single Nucleotide ,Article ,Databases ,03 medical and health sciences ,0302 clinical medicine ,Insulin resistance ,medicine ,Humans ,Precocious puberty ,SNP ,Allele ,Gene ,Data Management ,Genetics ,Multidisciplinary ,Chromosome ,medicine.disease ,Computational biology and bioinformatics ,030104 developmental biology ,Medicine ,Endocrine Cells ,Signal Transduction - Abstract
Precocious puberty (PP) is an important endocrine disorder affecting children globally. Several genes, SNPs and comorbidities are reported to be associated with PP; however, this data is scattered across scientific literature and has not been systematically collated and analysed. In this study, we present PrecocityDB as the first manually curated online database on genes and their ontology terms, SNPs, and pathways associated with PP. A tool for visualizing SNP coordinates and allelic variation on each chromosome, for genes associated with PP is also incorporated in PrecocityDB. Pathway enrichment analysis of PP-associated genes revealed that endocrine and cancer-related pathways are highly enriched. Disease enrichment analysis indicated that individuals with PP seem to be highly likely to suffer from reproductive and metabolic disorders such as PCOS, hypogonadism, and insulin resistance. PrecocityDB is a useful resource for identification of comorbid conditions and disease risks due to shared genes in PP. PrecocityDB is freely accessible at http://www.precocity.bicnirrh.res.in. The database source code and content can be downloaded through GitHub (https://github.com/bic-nirrh/precocity).
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- 2021
15. FSHR antagonists can trigger a PCOS-like state
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Faiza Hanif Waghu, Karishma Desai, Sumana Srinivasan, Kaushiki S. Prabhudesai, Vikas Dighe, Kareenhalli V. Venkatesh, and Susan Idicula-Thomas
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Phosphatidylinositol 3-Kinases ,endocrine system ,Reproductive Medicine ,Urology ,Animals ,Humans ,Receptors, FSH ,Female ,Testosterone ,Follicle Stimulating Hormone ,Luteinizing Hormone ,Polycystic Ovary Syndrome ,Rats - Abstract
Over the recent years, FSHR has become an important target for development of fertility regulating agents, as impairment of FSH-FSHR interaction can lead to subfertility or infertility. In our previous study, we identified a 9-mer peptide (FSHβ (89–97)) that exhibited FSHR antagonist activity. The histopathological and biochemical observations indicated, in addition to FSHR antagonism, a striking resemblance to a PCOS-like state. These observations led us to hypothesize that use of FSHR antagonists can trigger a PCOS-like state. In the present study, to validate this hypothesis, we performed qRT-PCR validation using ovarian tissue samples from our previous study. Expression of three genes known to be differentially expressed in PCOS was evaluated and found to be similar to the PCOS state. To further test the hypothesis, theoretical simulations were carried out by using the human menstrual cycle model available in the literature. Model simulations for FSHR antagonism were indicative of increased testosterone levels, increased ratio of luteinizing hormone/follicle stimulating hormone, and stockpiling of secondary follicles, which are typical characteristics of PCOS. The findings of this study will be relevant while reviewing the utility of FSHR antagonists for fertility regulation and reproductive medicine. Abbreviations: FSH: Follicle-stimulating hormone; FSHR: Follicle-stimulating hormone receptor; cAMP: Cyclic adenosine 3’5’ monophosphate; PKA: Protein kinase A; PI3K: Phosphoinositide 3-kinase; PKB: protein kinase B; ERK1/2: Extracellular signal-regulated protein kinase 1/2; MAPK: Mitogen-activated protein kinases; T: testosterone; E2: estradiol; PCOS: Polycystic ovarian syndrome; LH: luteinizing hormone; Lhcgr: luteinizing hormone/choriogonadotropin receptor; CYP17A1: cytochrome P450 family 17 subfamily A member 1; Inhba: inhibin subunit beta A; qRT-PCR: Real-Time quantitative reverse transcription polymerase chain reaction; FSHβ: Follicle-stimulating hormone β subunit; Ct: Cycle threshold; Rn18s: Rattus norvegicus 18S ribosomal RNA
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- 2021
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16. Membrane Interactome of a Recombinant Fragment of Human Surfactant Protein D Reveals GRP78 as a Novel Binding Partner in PC3, a Metastatic Prostate Cancer Cell Line
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Gargi Thakur, Gajanan Sathe, Indra Kundu, Barnali Biswas, Poonam Gautam, Saad Alkahtani, Susan Idicula-Thomas, Ravi Sirdeshmukh, Uday Kishore, and Taruna Madan
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0301 basic medicine ,lcsh:Immunologic diseases. Allergy ,Male ,GRP78 ,surfactant protein D ,medicine.drug_class ,Immunology ,Collectin ,Monoclonal antibody ,Interactome ,Metastasis ,law.invention ,03 medical and health sciences ,Prostate cancer ,0302 clinical medicine ,law ,medicine ,Immunology and Allergy ,Humans ,Neoplasm Metastasis ,Endoplasmic Reticulum Chaperone BiP ,innate immunity ,Heat-Shock Proteins ,Original Research ,Chemistry ,Surfactant protein D ,Cell Membrane ,apoptosis ,Prostatic Neoplasms ,medicine.disease ,Pulmonary Surfactant-Associated Protein D ,prostate cancer ,Recombinant Proteins ,Cell biology ,Neoplasm Proteins ,030104 developmental biology ,030220 oncology & carcinogenesis ,Cancer cell ,PC-3 Cells ,Recombinant DNA ,interactome analysis ,lcsh:RC581-607 ,signaling - Abstract
Data Availability Statement: The datasets presented in this study can be found in online repositories. The names of the repositories and accession number(s) can be found in the article/Supplementary Material (https://www.frontiersin.org/articles/10.3389/fimmu.2020.600660/full#h11). Copyright © 2021 Thakur, Sathe, Kundu, Biswas, Gautam, Alkahtani, Idicula-Thomas, Sirdeshmukh, Kishore and Madan. Surfactant protein-D (SP-D), a member of the collectin family has been shown to induce apoptosis in cancer cells. SP-D is composed of an N-terminal collagen-like domain and a calcium-dependent carbohydrate recognition domain (CRD). Recently, we reported that a recombinant fragment of human SP-D (rfhSP-D), composed of homotrimeric CRD region, induced intrinsic apoptotic pathway in prostate cancer cells. Here, we analyzed the membrane interactome of rfhSP-D in an androgen-independent prostate cancer cell line, PC3, by high resolution mass spectrometry and identified 347 proteins. Computational analysis of PPI network of this interactome in the context of prostate cancer metastasis and apoptosis revealed Glucose Regulated Protein of 78 kDa (GRP78) as an important binding partner of rfhSP-D. Docking studies suggested that rfhSP-D (CRD) bound to the substrate-binding domain of glycosylated GRP78. This was further supported by the observations that human recombinant GRP78 interfered with the binding of rfhSP-D to anti-SP-D polyclonal antibodies; GRP78 also significantly inhibited the binding of recombinant full-length human SP-D with a monoclonal antibody specific to the CRD in a dose-dependent manner. We conclude that the interaction with rfhSP-D is likely to interfere with the pro-survival signaling of GRP78. ICMR- National Institute for Research in Reproductive Health ICMR-NIRRH (Accession no. 921); ICMR- National Institute for Research in Reproductive Health ICMR-NIRRH-JRF; ICMR- National Institute for Research in Reproductive Health ICMR-SRF; Researchers Supporting Project (RSP-2020/26) King Saud University, Riyadh.
- Published
- 2020
17. A 5-mer peptide derived from hinge region of hFSHR can function as positive allosteric modulator in vivo
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Susan Idicula-Thomas, Vikas Dighe, Kaushiki S. Prabhudesai, and Muthu Sankar Aathi
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0301 basic medicine ,endocrine system ,Allosteric modulator ,Biophysics ,030209 endocrinology & metabolism ,Peptide ,Leucine-rich repeat ,Molecular Dynamics Simulation ,Biochemistry ,Second Messenger Systems ,Rats, Sprague-Dawley ,03 medical and health sciences ,0302 clinical medicine ,Allosteric Regulation ,Extracellular ,Cyclic AMP ,Animals ,Humans ,Receptor ,Granulosa cell proliferation ,Cell Proliferation ,chemistry.chemical_classification ,Granulosa Cells ,Chemistry ,Cell Biology ,Amino acid ,Cell biology ,Rats ,Molecular Docking Simulation ,Transmembrane domain ,030104 developmental biology ,HEK293 Cells ,Receptors, FSH ,Female ,Peptides - Abstract
Interaction of follicle stimulating hormone (FSH) with its cognate receptor (FSHR) is critical for maintaining reproductive health. FSHR has a large extracellular domain (ECD), composed of leucine rich repeats (LRRs) and hinge region, a transmembrane domain (TMD) and a short C-terminal domain (CTD). In this study, we have identified a short peptidic stretch in the hinge region (hFSHR(271-275)), through extensive computational modeling, docking and MD simulations, that is capable of independently interacting with the extracellular loops of FSHR(TMD). In vitro studies revealed that FSHR(271-275) peptide increased binding of [125I]-FSH to rat Fshr as well as FSH-induced cAMP production. Administration of FSHR(271-275) peptide in immature female rats significantly increased FSH-mediated ovarian weight gain and promoted granulosa cell proliferation. In summary, the results demonstrate that the synthetic peptide corresponding to amino acids 271-275 of hFSHR-hinge region stimulates FSH-FSHR interaction and behaves as positive allosteric modulator of FSHR. The study also lends evidence to the existing proposition that hinge region maintains the receptor in an inactive conformation in the absence of its ligand by engaging in intramolecular interactions with extracellular loops of TMD.
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- 2020
18. PCOSKB
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Mridula, Sharma, Ram Shankar, Barai, Indra, Kundu, Sameeksha, Bhaye, Khushal, Pokar, and Susan, Idicula-Thomas
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MicroRNAs ,Gene Expression Profiling ,Knowledge Bases ,Databases, Genetic ,Computational Biology ,Humans ,Disease ,Female ,Gene Regulatory Networks ,Polymorphism, Single Nucleotide ,Algorithms ,Polycystic Ovary Syndrome ,Signal Transduction - Abstract
PolyCystic Ovary Syndrome KnowledgeBase (PCOSKB
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- 2020
19. A QSAR modeling approach for predicting myeloid antimicrobial peptides with high sequence similarity
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Ulka Gawde, Evans C. Coutinho, Anish Narasimhan Gomatam, Faiza Hanif Waghu, and Susan Idicula-Thomas
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Models, Molecular ,Pore Forming Cytotoxic Proteins ,Quantitative structure–activity relationship ,Antimicrobial peptides ,Quantitative Structure-Activity Relationship ,Peptide ,Computational biology ,01 natural sciences ,Biochemistry ,Hemolysis ,Structure-Activity Relationship ,Similarity (network science) ,Drug Discovery ,Escherichia coli ,Humans ,Amino Acid Sequence ,Pharmacology ,chemistry.chemical_classification ,010405 organic chemistry ,Chemistry ,Organic Chemistry ,Rational design ,Reproducibility of Results ,Antimicrobial ,0104 chemical sciences ,Cathelicidins ,010404 medicinal & biomolecular chemistry ,Klebsiella pneumoniae ,Cheminformatics ,Pseudomonas aeruginosa ,Molecular Medicine - Abstract
Microbial resistance to conventional antibiotics has led to a surge in antimicrobial peptide (AMP) rational design initiatives that rely heavily on algorithms with good prediction accuracy and sensitivity. We present a quantitative structure-activity relationship (QSAR) approach for predicting activity of cathelicidins, an AMP family with broad-spectrum activity. The best multiple linear regression model built against Escherichia coli ATCC 25922 could accurately predict activity of three rationally designed peptides CP, DP, and Mapcon, having high sequence similarity. On further experimental validation of the rationally designed peptides, CP was found to exhibit high antimicrobial activity with negligible hemolysis. Here, we provide CP, an AMP with potential therapeutic applications and a family-based QSAR model for AMP prediction.
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- 2019
20. Tubulin acetylation: A novel functional avenue for CDYL in sperm
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Veena Dalvi, Shobha Sonawane, Susan Idicula-Thomas, Priyanka Parte, Sushma Mylavaram, Sweta Parab, and Abhipriya Kishore
- Subjects
Male ,0301 basic medicine ,Axoneme ,macromolecular substances ,Flagellum ,Microtubules ,Chromodomain ,Rats, Sprague-Dawley ,03 medical and health sciences ,Tubulin ,Structural Biology ,Microtubule ,Animals ,030102 biochemistry & molecular biology ,biology ,Acetylation ,Cell Biology ,Lysine Acetyltransferases ,Sperm ,Rats ,Molecular Docking Simulation ,030104 developmental biology ,Biochemistry ,Sperm Tail ,Acetyltransferase ,biology.protein - Abstract
Motility in sperm is driven by the flagella, the principal component of which is the axoneme. The microtubules which make up the 9 + 2 axoneme are composed of heterodimers of alpha and beta tubulins and undergo several post-translational modifications. We have earlier reported that HDAC6 functions as tubulin deacetylase in sperm and has a role in sperm movement. While exploring the specific tubulin acetyltransferase (TAT) in sperm we observed the presence of Chromodomain Y-Like (CDYL), on the principal piece of rat spermatozoa which compelled us to explore its function in sperm. CDYL was observed to be colocalized with acetylated alpha-tubulin (Ac α Tubulin) in sperm flagella. Sperm axonemal fraction showed the presence of CDYL protein indicating its strong association with flagellar microtubules. Sequence alignment of CDYL chromo domain and Alpha tubulin acetyltransferase (αTAT1) revealed that of the 10 residues of αTAT1 known to be involved in α-tubulin binding, 5 residues were identical and 1 was conserved between the two proteins. Docking of CDYL chromo domain and α-tubulin showed that 6 of the 11 important binding residues of α-tubulin showed an interaction with CDYL chromo domain. The putative CDYL chromodomain –α-tubulin interaction was further confirmed by Microscale Thermophoresis. We further asserted the ability of recombinant CDYL and Sperm CDYL to acetylate soluble tubulin and microtubules in vitro. Acetylation of tubulin was increased over two fold in cells overexpressing CDYL. Thus our studies convincingly demonstrate the ability of CDYL to moonlight as a tubulin acetyltransferase. This article is protected by copyright. All rights reserved.
- Published
- 2017
21. A steady-state modeling approach for simulation of antimicrobial peptide-cell membrane interaction
- Author
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Kareenhalli V. Venkatesh, Susan Idicula-Thomas, Sumana Srinivasan, and Faiza Hanif Waghu
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Models, Molecular ,Antimicrobial peptides ,Biophysics ,Peptide ,Cooperativity ,AMP binding ,Microbial Sensitivity Tests ,Biochemistry ,Cell membrane ,03 medical and health sciences ,medicine ,030304 developmental biology ,chemistry.chemical_classification ,0303 health sciences ,030306 microbiology ,Systems Biology ,Cell Membrane ,Membrane Proteins ,Cell Biology ,Anti-Bacterial Agents ,Membrane ,medicine.anatomical_structure ,chemistry ,Steady state (chemistry) ,Saturation (chemistry) ,Antimicrobial Cationic Peptides ,Protein Binding - Abstract
Antimicrobial Peptides (AMPs) are host defense molecules that initiate microbial death by binding to the membrane. On membrane binding, AMPs undergo changes in conformation and aggregation state to enable killing action. Depending on the AMP and cell membrane characteristics, the nature of binding can be aggregating or non-aggregating, with high/low cooperativity, at single or multiple sites with high/low affinity leading to a unique killing action that needs to be studied individually. In the present study, a steady-state model that simulates AMP-membrane interaction was developed and was used to predict the mechanism of AMP binding. The predictions obtained from the model were validated with experimentally deciphered values available in literature. The model was further used to predict the mechanism for a set of designed AMPs with high sequence similarity to Myeloid Antimicrobial Peptide (MAP) family. Depending on the predicted mechanism, a unique half saturation constant and steepness of response (Hill coefficient) was obtained which was further validated with available data from literature. The model could reliably predict the mechanism, the half saturation constant and the Hill coefficient values. Further based on the analysis, it was observed that aggregation and oligomerization result in drastic killing action in a short range of peptide concentration owing to high Hill coefficient values. Mechanisms such as monomers binding at multiple sites with/without cooperativity result in antimicrobial activity at low half saturation constant though the killing action may not be steep. Thus, the methodology developed here can be used to develop hypothesis for studying AMP-membrane interaction mechanisms.
- Published
- 2019
22. Identification and in vivo validation of a 9-mer peptide derived from FSHβ with FSHR antagonist activity
- Author
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Alessandro Contini, Sahil Raje, Susan Idicula-Thomas, Ankita Dhamanaskar, Vikas Dighe, Kaushiki S. Prabhudesai, and Deepak Modi
- Subjects
endocrine system ,Physiology ,030209 endocrinology & metabolism ,Peptide ,Crystallography, X-Ray ,Biochemistry ,Homology (biology) ,Rats, Sprague-Dawley ,Structure-Activity Relationship ,03 medical and health sciences ,Cellular and Molecular Neuroscience ,0302 clinical medicine ,Endocrinology ,Ovarian Follicle ,In vivo ,Animals ,chemistry.chemical_classification ,Granulosa Cells ,Estradiol ,Chemistry ,Ovary ,Antagonist ,Cell cycle ,In vitro ,Rats ,Cell biology ,Follicle Stimulating Hormone, beta Subunit ,Receptors, FSH ,Female ,Protein Structural Elements ,Folliculogenesis ,Oligopeptides ,Spermatogenesis ,030217 neurology & neurosurgery - Abstract
FSH-FSHR interaction is critical for folliculogenesis, spermatogenesis and progression of several cancers. Therefore, FSHR is an attractive target for fertility regulation and cancer therapeutics. Based on homology and structural analysis of hFSH-FSHR(ECD) complex, a minimal continuous stretch within FSHβ seat-belt loop (FSHβ (89-97)) was identified to be crucial for FSHR interaction. The ability of FSHβ (89-97) peptide to neutralize FSHR activity was evaluated by a panel of in vitro and in vivo experiments. The synthetic peptide significantly inhibited binding of [125I]-FSH to rat Fshr as well as FSH-induced cAMP production. In immature rats, FSHβ (89-97) peptide administration reduced FSH-mediated increase in ovarian weight. The peptide inhibited transition of follicles from pre-antral to antral stage and hindered the cell cycle progression of granulosa cells beyond G0/G1 phase. In adult rats, administration of the peptide inhibited estradiol synthesis and significantly perturbed folliculogenesis.
- Published
- 2020
23. PCOSKB: A KnowledgeBase on genes, diseases, ontology terms and biochemical pathways associated with PolyCystic Ovary Syndrome
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Susan Idicula-Thomas, Ram Shankar Barai, Shaini Joseph, and Rasika Bhujbalrao
- Subjects
0301 basic medicine ,Databases, Factual ,Biochemical Phenomena ,Protein Conformation ,Knowledge Bases ,Gene regulatory network ,MEDLINE ,Single-nucleotide polymorphism ,Comorbidity ,Scientific literature ,Biology ,Bioinformatics ,Polymorphism, Single Nucleotide ,03 medical and health sciences ,Polymorphism (computer science) ,Genetics ,medicine ,Humans ,Database Issue ,Gene Regulatory Networks ,Gene ,medicine.disease ,Polycystic ovary ,Gene Ontology ,030104 developmental biology ,Mutation ,Female ,Databases, Chemical ,Polycystic Ovary Syndrome - Abstract
Polycystic ovary syndrome (PCOS) is one of the major causes of female subfertility worldwide and ≈ 7-10% of women in reproductive age are affected by it. The affected individuals exhibit varying types and levels of comorbid conditions, along with the classical PCOS symptoms. Extensive studies on PCOS across diverse ethnic populations have resulted in a plethora of information on dysregulated genes, gene polymorphisms and diseases linked to PCOS. However, efforts have not been taken to collate and link these data. Our group, for the first time, has compiled PCOS-related information available through scientific literature; cross-linked it with molecular, biochemical and clinical databases and presented it as a user-friendly, web-based online knowledgebase for the benefit of the scientific and clinical community. Manually curated information on associated genes, single nucleotide polymorphisms, diseases, gene ontology terms and pathways along with supporting reference literature has been collated and included in PCOSKB (http://pcoskb.bicnirrh.res.in).
- Published
- 2015
24. PBIT: Pipeline Builder for Identification of drug Targets for infectious diseases
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Mohammed Husain Bharmal, Vinit Shetty, Gauri Shende, Ram Shankar Barai, Susan Idicula-Thomas, and Harshala Haldankar
- Subjects
0301 basic medicine ,Statistics and Probability ,Drug ,030103 biophysics ,Proteome ,Computer science ,In silico ,media_common.quotation_subject ,Bioinformatics ,Communicable Diseases ,Biochemistry ,03 medical and health sciences ,Human gut ,Candida albicans ,Drug Discovery ,Human proteome project ,Humans ,Computer Simulation ,Molecular Biology ,media_common ,Supplementary data ,Internet ,Microbiota ,Computational Biology ,Pipeline (software) ,Computer Science Applications ,Computational Mathematics ,030104 developmental biology ,Computational Theory and Mathematics ,Identification (biology) - Abstract
Summary PBIT (Pipeline Builder for Identification of drug Targets) is an online webserver that has been developed for screening of microbial proteomes for critical features of human drug targets such as being non-homologous to human proteome as well as the human gut microbiota, essential for the pathogen’s survival, participation in pathogen-specific pathways etc. The tool has been validated by analyzing 57 putative targets of Candida albicans documented in literature. PBIT integrates various in silico approaches known for drug target identification and will facilitate high-throughput prediction of drug targets for infectious diseases, including multi-pathogenic infections. Availability and Implementation PBIT is freely accessible at http://www.pbit.bicnirrh.res.in/. Supplementary information Supplementary data are available at Bioinformatics online.
- Published
- 2016
25. Effect of diversity in gp41 membrane proximal external region of primary HIV-1 Indian subtype C sequences on interaction with broadly neutralizing antibodies 4E10 and 10E8
- Author
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Vidya S. Nagar, Priya Patil, Vainav Patel, Jyoti Sutar, Atmaram H. Bandivdekar, Jayanta Bhattacharya, Dhanashree D. Jagtap, Bhalachandra Kulkarni, Nitin Hingankar, Suprit Deshpande, Susan Idicula-Thomas, Archana Sonawani, and Varsha Padwal
- Subjects
Adult ,Male ,Cancer Research ,Human immunodeficiency virus (HIV) ,India ,HIV Infections ,HIV Antibodies ,Gp41 ,medicine.disease_cause ,Deep sequencing ,Neutralization ,DNA sequencing ,Epitope ,Epitopes ,03 medical and health sciences ,Meta-Analysis as Topic ,Neutralization Tests ,Virology ,medicine ,Humans ,Child ,030304 developmental biology ,0303 health sciences ,biology ,030306 microbiology ,Haplotype ,Genetic Variation ,High-Throughput Nucleotide Sequencing ,Middle Aged ,HIV Envelope Protein gp41 ,Molecular Docking Simulation ,Infectious Diseases ,HIV-1 ,biology.protein ,Female ,Antibody ,Broadly Neutralizing Antibodies - Abstract
Human Immunodeficiency Virus-1 Clade C (HIV-1C) dominates the AIDS epidemic in India, afflicting 2.1 million individuals within the country and more than 15 million people worldwide. Membrane proximal external region (MPER) is an attractive target for broadly neutralizing antibody (bNAb) based therapies. However, information on MPER sequence diversity from India is meagre due to limited sampling of primary viral sequences. In the present study, we examined the variation in MPER of HIV-1C from 24 individuals in Mumbai, India by high throughput sequencing of uncultured viral sequences. Deep sequencing of MPER (662-683; HXB2 envelope amino acid numbering) allowed quantification of intra-individual variation up to 65% at positions 662, 665, 668, 674 and 677 within this region. These variable positions included contact sites targeted by bNAbs 2F5, Z13e1, 4E10 as well as 10E8. Both major and minor epitope variants i.e. 'haplotypes' were generated for each sample dataset. A total of 23, 34 and 25 unique epitope haplotypes could be identified for bNAbs 2F5, Z13e1 and 4E10/10E8 respectively. Further analysis of 4E10 and 10E8 epitopes from our dataset and meta-analysis of previously reported HIV-1 sequences from India revealed 26 epitopes (7 India-specific), heretofore untested for neutralization sensitivity. Peptide-Ab docking predicted 13 of these to be non-binding to 10E8. ELISA, Surface Plasmon Resonance and peptide inhibition of HIV-1 neutralization assays were then performed which validated predicted weak/non-binding interactions for peptides corresponding to six of these epitopes. These results highlight the under-representation of 10E8 non-binding HIV-1C MPER sequences from India. Our study thus underscores the need for increased surveillance of primary circulating envelope sequences for development of efficacious bNAb-based interventions in India.
- Published
- 2019
26. Evaluation of 4-thiazolidinone derivatives as potential reverse transcriptase inhibitors against HIV-1 drug resistant strains
- Author
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Srikanth Tripathy, Dipen Desai, Ramesh S. Paranjape, Smita Kulkarni, Nandini Makwana, Vanangamudi Murugesan, Seturam B. Katti, Rahul K. Suryawanshi, Devidas N. Chaturbhuj, Susan Idicula-Thomas, Archana Sonawani, and Sushama Jadhav
- Subjects
Nevirapine ,Anti-HIV Agents ,In silico ,Human immunodeficiency virus (HIV) ,HIV Infections ,Drug resistance ,medicine.disease_cause ,01 natural sciences ,Biochemistry ,Cell Line ,Structure-Activity Relationship ,Drug Discovery ,Drug Resistance, Viral ,medicine ,Humans ,Molecular Biology ,IC50 ,Strain (chemistry) ,010405 organic chemistry ,Chemistry ,Organic Chemistry ,Virology ,Reverse transcriptase ,In vitro ,HIV Reverse Transcriptase ,0104 chemical sciences ,Molecular Docking Simulation ,010404 medicinal & biomolecular chemistry ,HIV-1 ,Leukocytes, Mononuclear ,Reverse Transcriptase Inhibitors ,Thiazolidines ,medicine.drug - Abstract
Rapid emergence of drug resistance is crucial in management of HIV infection limiting implementation of efficacious drugs in the ART regimen. Designing new molecules against HIV drug resistant strains is utmost essential. Based on the anti-HIV-1 activity, we selected four 4-thiazolidinone derivatives (S009-1908, S009-1909, S009-1911, S009-1912) and studied their interaction with reverse transcriptase (RT) from a panel of 10 clinical isolates (8 nevirapine resistant and two susceptible) using in silico methods, and inhibition pattern using in vitro cell based assays. On the basis of binding affinity observed in in silico analysis, 2-(2-chloro-6-nitrophenyl)-3-(4, 6-dimethylpyridin-2-yl) thiazolidin-4-one (S009-1912) was identified as the lead molecule followed by S009-1908, S009-1909 and S009-1911. The in vitro activity against the same panel was assessed using TZM-bl assay (IC50: 0.4–11.44 µg/ml, TI: 4–126) and subsequently in PBMC assay against a nevirapine resistant clinical isolate (IC50: 0.8–6.65 µg/ml, TI: 8.31–11.43) and standard strain from NIH ARRRP (IC50: 0.95–3.6 µg/ml, TI: 9–26). The study shows analogue with pyrimidin-2-yl amino substitution at N-3 position of thiazolidin-4-one ring (S009-1908, S009-1909, S009-1911) exhibited enhanced activity as compared to pyridin-2-yl substituted derivatives (S009-1912), suggesting the use 4-thiazolidinones for developing potent inhibitors against HIV-1 drug resistant strains.
- Published
- 2017
27. Cervical cancer in Indian women reveals contrasting association among common sub-family of HLA class I alleles
- Author
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Jayanti Mania-Pramanik, Archana Sonawani, Vinita Salvi, Hemangi Chaudhari, Priyanka Gokhale, Hemant B. Tongaonkar, Shilpa Kerkar, Himangi Warke, and Susan Idicula-Thomas
- Subjects
Adult ,Models, Molecular ,Cervical swab (specimen) ,Protein Conformation ,Immunology ,India ,Uterine Cervical Neoplasms ,Human leukocyte antigen ,Alphapapillomavirus ,Biology ,White People ,Epitope ,Epitopes ,symbols.namesake ,Gene Frequency ,Genetics ,medicine ,Humans ,Genetic Predisposition to Disease ,Typing ,Allele ,Alleles ,Aged ,Cervical cancer ,Histocompatibility Antigens Class I ,Reproducibility of Results ,Middle Aged ,medicine.disease ,Human genetics ,Bonferroni correction ,Case-Control Studies ,symbols ,Female ,Protein Binding - Abstract
We studied the relationship between human leukocyte antigen (HLA) class I alleles and cervical cancer among Indian women. Seventy-five cervical cancer cases were compared with 175 noncancer controls. Cervical biopsy tissue specimen from cancer cases and cervical swab specimen from controls were collected for HPV detection and typing. Blood was taken for HLA typing by PCR-SSOP method. The impact of HLA class I alleles on cervical cancer risk was evaluated using StatCalc program (Epi Info version 6.0.4. CDC Atlanta, GA, USA), and confirmed with Bonferroni correction. Results revealed HLA-B*37, HLA-B*58 were associated significantly with increased risk while HLA-B*40 with decreased risk for cervical cancer. At high-resolution analysis after Bonferroni correction, HLA-B*37:01 allele was associated with increased risk, whereas HLA-B*40:06 was with decreased risk for cervical cancer. HLA-B*37:01 and HLA-B*40:06 belong to the same superfamily of HLA-B44. In silico analysis revealed different binding affinities of HLA-B*37:01 and HLA-B*40:06 for the epitopes predicted for E6 and L1 proteins of HPV16. The higher binding affinity of epitopes to B*40:06, as revealed by docking studies, supports the hypothesis that this allele is able to present the antigenic peptides more efficiently than B*37:01 and thereby can protect the carriers from the risk of cervical cancer. Thus, there is a clear indication that HLA plays an important role in the development of cervical cancer in HPV-infected women. Identification of these factors in high-risk HPV-infected women may help in reducing the cervical cancer burden in India.
- Published
- 2014
28. HPV16 E6 variants: Frequency, association with HPV types and in silico analysis of the identified novel variants
- Author
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Shilpa Kerkar, Priyanka Gokhale, Archana Sonawani, Hemant B. Tongaonkar, Jayanti Mania-Pramanik, and Susan Idicula-Thomas
- Subjects
Cervical cancer ,Genetics ,Lineage (genetic) ,Phylogenetic tree ,Hpv types ,Sequence analysis ,In silico ,Biology ,medicine.disease ,Virology ,Hpv16 e6 ,Infectious Diseases ,medicine ,In patient - Abstract
High-risk human papillomavirus (HPV) types, specifically HPV 16 E6 variants are involved in viral persistence and the development of cervical lesions. India contributes to 1/3rd of the global cervical cancer deaths; however, information on E6 variants in the Indian population is limited. Information on these variants is essential for successful implementation of cervical cancer immunization programs. The E6 variants and their possible biological implications to the outcome of infection were studied in women attending the Tata Memorial Hospital, Mumbai, India. Cervical cancer patients with HPV 16 as a single infection (n = 33), co-infection with another HPV type (n = 20) or with multiple types (n = 10) were examined for HPV16 E6 variants using PCR and sequence analysis. The variants were identified using the prototype sequence (HPV 16R) belonging to the European lineage. The results revealed that the European T350G was the most common variant (50%) followed by the European prototype (40.3%) and the North-American (N = 3; 4.8%). The European prototype was significantly more frequent in patients infected with HPV16 alone (P
- Published
- 2014
29. CAMP: Collection of sequences and structures of antimicrobial peptides
- Author
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Faiza Hanif Waghu, Susan Idicula-Thomas, Ram Shankar Barai, Lijin Gopi, Bilal Nizami, and Pranay Ramteke
- Subjects
Internet ,Structure analysis ,Sequence analysis ,Antimicrobial peptides ,Molecular Conformation ,Computational biology ,Biology ,Antimicrobial ,Molecular conformation ,Microbiology ,VI. Genomic variation, diseases and drugs ,Anti-Infective Agents ,Sequence Analysis, Protein ,Genetics ,Peptides ,Algorithms ,Databases, Chemical - Abstract
Antimicrobial peptides (AMPs) are gaining importance as anti-infective agents. Here we describe the updated Collection of Antimicrobial Peptide (CAMP) database, available online at http://www.camp.bicnirrh.res.in/. The 3D structures of peptides are known to influence antimicrobial activity. Although there exists databases of AMPs, information on structures of AMPs is limited in these databases. CAMP is manually curated and currently holds 6756 sequences and 682 3D structures of AMPs. Sequence and structure analysis tools have been incorporated to enhance the usefulness of the database.
- Published
- 2013
30. A Chemical Proteomics Approach to Profiling the ATP-binding Proteome of Mycobacterium tuberculosis
- Author
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Stephan C. Schürer, Lisa M. Wolfe, Susan Idicula-Thomas, Karen M. Dobos, Krister Wennerberg, Robert C. Reynolds, Gurdyal S. Besra, and Usha Veeraraghavan
- Subjects
Proteomics ,Tuberculosis ,Proteome ,Antitubercular Agents ,Biology ,Binding, Competitive ,Biochemistry ,Analytical Chemistry ,Mycobacterium tuberculosis ,03 medical and health sciences ,Adenosine Triphosphate ,Bacterial Proteins ,Lipid biosynthesis ,Protein Interaction Mapping ,medicine ,Protein Isoforms ,Molecular Biology ,030304 developmental biology ,chemistry.chemical_classification ,0303 health sciences ,030306 microbiology ,Research ,Gene Expression Regulation, Bacterial ,biology.organism_classification ,medicine.disease ,Isocitrate Lyase ,Culture Media ,3. Good health ,DNA-Binding Proteins ,Oxygen ,Metabolic pathway ,Enzyme ,Regulon ,chemistry ,Carrier Proteins ,Peptides ,Protein Kinases ,Protein Binding ,Signal Transduction - Abstract
Tuberculosis, caused by Mycobacterium tuberculosis, remains one of the leading causes of death worldwide despite extensive research, directly observed therapy using multidrug regimens, and the widespread use of a vaccine. The majority of patients harbor the bacterium in a state of metabolic dormancy. New drugs with novel modes of action are needed to target essential metabolic pathways in M. tuberculosis; ATP-competitive enzyme inhibitors are one such class. Previous screening efforts for ATP-competitive enzyme inhibitors identified several classes of lead compounds that demonstrated potent anti-mycobacterial efficacy as well as tolerable levels of toxicity in cell culture. In this report, a probe-based chemoproteomic approach was used to selectively profile the M. tuberculosis ATP-binding proteome in normally growing and hypoxic M. tuberculosis. From these studies, 122 ATP-binding proteins were identified in either metabolic state, and roughly 60% of these are reported to be essential for survival in vitro. These data are available through ProteomeXchange with identifier PXD000141. Protein families vital to the survival of the tubercle bacillus during hypoxia emerged from our studies. Specifically, along with members of the DosR regulon, several proteins involved in energy metabolism (Icl/Rv0468 and Mdh/Rv1240) and lipid biosynthesis (UmaA/Rv0469, DesA1/Rv0824c, and DesA2/Rv1094) were found to be differentially abundant in hypoxic versus normal growing cultures. These pathways represent a subset of proteins that may be relevant therapeutic targets for development of novel ATP-competitive antibiotics.
- Published
- 2013
31. Leveraging family-specific signatures for AMP discovery and high-throughput annotation
- Author
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Ram Shankar Barai, Susan Idicula-Thomas, and Faiza Hanif Waghu
- Subjects
0301 basic medicine ,Computer science ,medicine.drug_class ,Antibiotics ,Antimicrobial peptides ,Computational biology ,computer.software_genre ,medicine.disease_cause ,Article ,03 medical and health sciences ,Annotation ,Microbial resistance ,medicine ,Amino Acid Sequence ,Escherichia coli ,Peptide sequence ,Throughput (business) ,Multidisciplinary ,Sequence Homology, Amino Acid ,030102 biochemistry & molecular biology ,Rational design ,Biological activity ,Antimicrobial ,High-Throughput Screening Assays ,030104 developmental biology ,Data mining ,computer ,Antimicrobial Cationic Peptides - Abstract
Antimicrobial peptides (AMPs) are diverse, biologically active, essential components of the innate immune system. As compared to conventional antibiotics, AMPs exhibit broad spectrum antimicrobial activity, reduced toxicity and reduced microbial resistance. They are widely researched for their therapeutic potential, especially against multi-drug resistant pathogens. AMPs are known to have family-specific sequence composition, which can be mined for their discovery and rational design. Here, we present a detailed family-based study on AMP families. The study involved the use of sequence signatures represented by patterns and hidden Markov models (HMMs) present in experimentally studied AMPs to identify novel AMPs. Along with AMPs, peptides hitherto lacking antimicrobial annotation were also retrieved and wet-lab studies on randomly selected sequences proved their antimicrobial activity against Escherichia coli. CAMPSign, a webserver has been created for researchers to effortlessly exploit the use of AMP family signatures for identification of AMPs. The webserver is available online at www.campsign.bicnirrh.res.in. In this work, we demonstrate an optimised and experimentally validated protocol along with a freely available webserver that uses family-based sequence signatures for accelerated discovery of novel AMPs.
- Published
- 2016
32. ClassAMP: A Prediction Tool for Classification of Antimicrobial Peptides
- Author
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Susan Idicula-Thomas, Shreyas Karnik, Shaini Joseph, Pravin Nilawe, and V. K. Jayaraman
- Subjects
Antifungal ,Antiinfective agent ,Support Vector Machine ,Drug discovery ,medicine.drug_class ,Applied Mathematics ,Antimicrobial peptides ,Computational biology ,Biology ,Bioinformatics ,Antimicrobial ,Random forest ,Support vector machine ,Protein sequencing ,Anti-Infective Agents ,Genetics ,medicine ,Peptides ,Algorithms ,Biotechnology - Abstract
Antimicrobial peptides (AMPs) are gaining popularity as anti-infective agents. Information on sequence features that contribute to target specificity of AMPs will aid in accelerating drug discovery programs involving them. In this study, an algorithm called ClassAMP using Random Forests (RFs) and Support Vector Machines (SVMs) has been developed to predict the propensity of a protein sequence to have antibacterial, antifungal, or antiviral activity. ClassAMP is available at http://www.bicnirrh.res.in/classamp/.
- Published
- 2012
33. Novel homozygous mutations in Desert Hedgehog gene in patients with 46,XY complete gonadal dysgenesis and prediction of its structural and functional implications by computational methods
- Author
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Susan Idicula-Thomas, Dhanjit Kumar Das, Daksha Sanghavi, Lakshmi Vasudevan, and Harshavardhan M. Gawde
- Subjects
Adult ,Models, Molecular ,Adolescent ,DNA Mutational Analysis ,Molecular Sequence Data ,Mutant ,India ,Gonadal dysgenesis ,Biology ,medicine.disease_cause ,Polymerase Chain Reaction ,XY gonadal dysgenesis ,Exon ,Gene duplication ,Genetics ,medicine ,Humans ,Hedgehog Proteins ,Amino Acid Sequence ,Genes, sry ,Genetics (clinical) ,Desert hedgehog ,Sequence Deletion ,Gonadal Dysgenesis, 46,XY ,Mutation ,Chromosomes, Human, Pair 12 ,Base Sequence ,Homozygote ,Exons ,General Medicine ,medicine.disease ,Testis determining factor ,Karyotyping ,Female ,Protein Binding - Abstract
Male to female sex reversal in patients with 46,XY karyotype results from the failure of development of testis which may be due to mutations in the SRY gene. Only 10-15% of cases of 46,XY gonadal dysgenesis are accounted for by different types of mutations in the SRY gene. Hence, majority of such patients may have mutations in other genes involved in the testicular differentiation pathway. Besides SRY, other autosomal and X-linked genes are also involved in sexual development during embryogenesis. We describe here the first report from India wherein, two cases of 46,XY complete gonadal dysgenesis that could be attributable to mutations in the Desert hedgehog (DHH) gene. The mutations found in these two patients were a homozygous deletion (c.271_273delGAG) that resulted in deletion of one amino acid (p.D90del) and a homozygous duplication (c.57-60dupAGCC) that resulted in premature termination resulting in non-functional DHH protein. The structure-function implications of the p.D90del mutation were predicted using computational tools. Structural studies on the p.D90del mutant revealed that the mutation could seriously perturb the interaction of DHH with its binding partners. This is the second report in literature showing homozygous mutation in cases with 46,XY complete gonadal dysgenesis.
- Published
- 2011
34. CAMP: a useful resource for research on antimicrobial peptides
- Author
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V. K. Jayaraman, Ram Shankar Barai, Shaini Thomas, Susan Idicula-Thomas, and Shreyas Karnik
- Subjects
Sequence analysis ,Antimicrobial peptides ,Information Storage and Retrieval ,Computational biology ,Biology ,Bioinformatics ,Anti-Infective Agents ,Databases, Genetic ,Genetics ,Humans ,Databases, Protein ,Internet ,Online database ,Computational Biology ,Reproducibility of Results ,Articles ,Resource (Windows) ,Antimicrobial ,Random forest ,Support vector machine ,Mic values ,Databases, Nucleic Acid ,Peptides ,Algorithms ,Genome, Bacterial ,Software ,Antimicrobial Cationic Peptides - Abstract
Antimicrobial peptides (AMPs) are gaining popularity as better substitute to antibiotics. These peptides are shown to be active against several bacteria, fungi, viruses, protozoa and cancerous cells. Understanding the role of primary structure of AMPs in their specificity and activity is essential for their rational design as drugs. Collection of Anti-Microbial Peptides (CAMP) is a free online database that has been developed for advancement of the present understanding on antimicrobial peptides. It is manually curated and currently holds 3782 antimicrobial sequences. These sequences are divided into experimentally validated (patents and non-patents: 2766) and predicted (1016) datasets based on their reference literature. Information like source organism, activity (MIC values), reference literature, target and non-target organisms of AMPs are captured in the database. The experimentally validated dataset has been further used to develop prediction tools for AMPs based on the machine learning algorithms like Random Forests (RF), Support Vector Machines (SVM) and Discriminant Analysis (DA). The prediction models gave accuracies of 93.2% (RF), 91.5% (SVM) and 87.5% (DA) on the test datasets. The prediction and sequence analysis tools, including BLAST, are integrated in the database. CAMP will be a useful database for study of sequence-activity and -specificity relationships in AMPs. CAMP is freely available at http://www.bicnirrh.res.in/antimicrobial.
- Published
- 2009
35. Phenotypic and genotypic characterization of Factor VII deficiency patients from Western India
- Author
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Shrimati Shetty, Kanjaksha Ghosh, Leenam Mota, and Susan Idicula-Thomas
- Subjects
Adult ,Male ,Models, Molecular ,Adolescent ,Protein Conformation ,Factor VII Deficiency ,Molecular Sequence Data ,Clinical Biochemistry ,Mutant ,India ,medicine.disease_cause ,Biochemistry ,Conserved sequence ,Young Adult ,chemistry.chemical_compound ,Asian People ,hemic and lymphatic diseases ,Genotype ,medicine ,Humans ,Missense mutation ,Amino Acid Sequence ,cardiovascular diseases ,Child ,Blood Coagulation ,Gene ,Conserved Sequence ,Genetics ,Serine protease ,Mutation ,Polymorphism, Genetic ,biology ,Factor VII ,Protein Stability ,Biochemistry (medical) ,Infant ,General Medicine ,Middle Aged ,Phenotype ,Haplotypes ,chemistry ,Child, Preschool ,biology.protein ,Female - Abstract
Background Congenital factor VII (FVII) deficiency is a rare coagulation deficiency caused due to defects in the FVII gene. Methods We analyzed 14 unrelated Indian patients with congenital FVII deficiency for mutations in FVII gene by conformation sensitive gel electrophoresis (CSGE) followed by DNA sequencing. Results A total of 11 different missense mutations were identified, of which 5 were novel (Ala191Pro, Asp338Glu, Ile138Thr, Leu263Arg and Trp284Arg) and 6 had been previously reported (Cys22Arg, Arg152Gln, Cys310Phe, Thr324Met, Gly117Arg and His348Arg). Six of the 11 mutations were located in the catalytic serine protease domain, 3 in the activation domain and 1 each in the Gla and the second epidermal growth factor domain respectively. Multiple sequence alignment using ClustalW2 analysis showed that all the mutations were found in residues that are highly conserved across species. Implications of mutations on the structural stability and function of human factor VIII (hFVII) using Swiss-Pdb Viewer and the intra-molecular interactions of the mutant residues using PIC showed that there is a structural instability in all the mutants either by steric hindrance or instability in the protein molecule folding. Conclusion A wide spectrum of mutations was detected in the FVII gene; the presence of 6 out of 11 mutations in the serine protease domain suggests the crucial role of catalytic domain in FVII functional activity.
- Published
- 2009
36. A support vector machine-based method for predicting the propensity of a protein to be soluble or to form inclusion body on overexpression in Escherichia coli
- Author
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Abhijit Kulkarni, Petety V. Balaji, Bhaskar D. Kulkarni, Susan Idicula-Thomas, and Valadi K. Jayaraman
- Subjects
Statistics and Probability ,Directed Evolution ,Eukaryotic Proteins ,Mutant ,Computational biology ,Biology ,medicine.disease_cause ,Biochemistry ,Inclusion bodies ,Pattern Recognition, Automated ,Artificial Intelligence ,Sequence Analysis, Protein ,Catalytic Domain ,Escherichia coli ,medicine ,Fold Recognition ,Databases, Protein ,Molecular Biology ,Inclusion Bodies ,chemistry.chemical_classification ,Escherichia coli Proteins ,Gene Expression Profiling ,Point mutation ,A protein ,Gene Expression Regulation, Bacterial ,Classification ,Recombinant Proteins ,Computer Science Applications ,Amino acid ,Support vector machine ,Computational Mathematics ,Solubility ,Computational Theory and Mathematics ,chemistry ,Structural Genomics ,Protein folding ,Mutations ,Algorithms ,Expression Data - Abstract
Motivation: Inclusion body formation has been a major deterrent for overexpression studies since a large number of proteins form insoluble inclusion bodies when overexpressed in Escherichia coli. The formation of inclusion bodies is known to be an outcome of improper protein folding; thus the composition and arrangement of amino acids in the proteins would be a major influencing factor in deciding its aggregation propensity. There is a significant need for a prediction algorithm that would enable the rational identification of both mutants and also the ideal protein candidates for mutations that would confer higher solubility-on-overexpression instead of the presently used trial-and-error procedures. Results: Six physicochemical properties together with residue and dipeptide-compositions have been used to develop a support vector machine-based classifier to predict the overexpression status in E.coli. The prediction accuracy is ∼72% suggesting that it performs reasonably well in predicting the propensity of a protein to be soluble or to form inclusion bodies. The algorithm could also correctly predict the change in solubility for most of the point mutations reported in literature. This algorithm can be a useful tool in screening protein libraries to identify soluble variants of proteins. Avalibility: Software is available on request from the authors. Contact: balaji@iitcb.ac.in; vk.jayaraman@ncl.res.in Supplementary information: Supplementary data are available at Bioinformatics Online web site.
- Published
- 2005
37. Understanding the relationship between the primary structure of proteins and its propensity to be soluble on overexpression inEscherichia coli
- Author
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Petety V. Balaji and Susan Idicula-Thomas
- Subjects
chemistry.chemical_classification ,Hot Temperature ,Chemistry ,Protein primary structure ,Tripeptide ,medicine.disease_cause ,Biochemistry ,Protein Structure, Secondary ,Recombinant Proteins ,Article ,Inclusion bodies ,Amino acid ,Molecular Weight ,Protein structure ,Solubility ,Escherichia coli ,medicine ,Point Mutation ,Amino Acid Sequence ,Cloning, Molecular ,Molecular Biology ,Peptide sequence - Abstract
Solubility of proteins on overexpression in Escherichia coli is a manifestation of the net effect of several sequence-dependent and sequence-independent factors. This study aims to delineate the relationship between the primary structure and solubility on overexpression. The amino acid sequences of proteins reported to be soluble or to form inclusion bodies on overexpression in E. coli under normal growth conditions were analyzed. The results show a positive correlation between thermostability and solubility of proteins, and an inverse correlation between the in vivo half-life of proteins and solubility. The amino acid (Asn, Thr, Tyr) composition and the tripeptide frequency of the protein were also found to influence its solubility on overexpression. The amino acids that were seen to be present in a comparatively higher frequency in inclusion body-forming proteins have a higher sheet propensity, whereas those that are seen more in soluble proteins have a higher helix propensity; this is indicative of a possible correlation between sheet propensity and inclusion body formation. Thus, the present analysis shows that thermostability, in vivo half-life, Asn, Thr, and Tyr content, and tripeptide composition of a protein are correlated to the propensity of a protein to be soluble on overexpression in E. coli. The precise mechanism by which these properties affect the solubility status of the overexpressed protein remains to be understood.
- Published
- 2005
38. Imatinib resistance due to a novel tri-nucleotide insertion in the SH2/C lobe interface of BCR/ABL kinase domain between position K357 and N358
- Author
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Babu Rao Vundinti, Kanjaksha Ghosh, Susan Idicula-Thomas, Shantashri Vaidya, and Archana Sonawani
- Subjects
chemistry.chemical_classification ,Cancer Research ,Oncology ,chemistry ,Imatinib resistance ,Domain (ring theory) ,Cancer research ,Nucleotide ,Hematology ,Bcr abl kinase - Published
- 2012
39. HPV16 E6 variants: frequency, association with HPV types and in silico analysis of the identified novel variants
- Author
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Priyanka S, Gokhale, Archana, Sonawani, Susan, Idicula-Thomas, Shilpa, Kerkar, Hemant, Tongaonkar, and Jayanti, Mania-Pramanik
- Subjects
Adult ,Models, Molecular ,Human papillomavirus 16 ,Molecular Epidemiology ,Genotype ,Protein Conformation ,Papillomavirus Infections ,Genetic Variation ,India ,Uterine Cervical Neoplasms ,Oncogene Proteins, Viral ,Sequence Analysis, DNA ,Middle Aged ,Polymerase Chain Reaction ,Molecular Docking Simulation ,Repressor Proteins ,DNA, Viral ,Prevalence ,Humans ,Female ,Aged - Abstract
High-risk human papillomavirus (HPV) types, specifically HPV 16 E6 variants are involved in viral persistence and the development of cervical lesions. India contributes to 1/3rd of the global cervical cancer deaths; however, information on E6 variants in the Indian population is limited. Information on these variants is essential for successful implementation of cervical cancer immunization programs. The E6 variants and their possible biological implications to the outcome of infection were studied in women attending the Tata Memorial Hospital, Mumbai, India. Cervical cancer patients with HPV 16 as a single infection (n = 33), co-infection with another HPV type (n = 20) or with multiple types (n = 10) were examined for HPV16 E6 variants using PCR and sequence analysis. The variants were identified using the prototype sequence (HPV 16R) belonging to the European lineage. The results revealed that the European T350G was the most common variant (50%) followed by the European prototype (40.3%) and the North-American (N = 3; 4.8%). The European prototype was significantly more frequent in patients infected with HPV16 alone (P 0.05, C.I. 1.2-13.6), while the European T350G variants were seen in women with co-infections. The North-American lineage was found in women infected with HPV16 and 33. Three novel variants were identified of which two were non-synonymous. Phylogenetic analysis revealed that the variant F69L + L83V is not related to any of these lineages, while the variant M137L + L83V is closely related to the North American variant. This study found a difference in the prevalence of E6 variants compared to earlier Indian studies and their association with type of infection.
- Published
- 2014
40. Hereditary basis of protein C deficiency (PCD) in thrombosis patients: First report from India
- Author
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Susan Idicula-Thomas, Bipin Kulkarni, Kanjaksha Ghosh, Navin Pai, and Shrimati Shetty
- Subjects
Adult ,Male ,Pathology ,medicine.medical_specialty ,animal structures ,Adolescent ,DNA Mutational Analysis ,Mutation, Missense ,India ,Frameshift mutation ,Risk Factors ,Protein C deficiency ,otorhinolaryngologic diseases ,Coagulopathy ,medicine ,Humans ,Missense mutation ,Genetic Predisposition to Disease ,Frameshift Mutation ,Blood Coagulation ,Venous Thrombosis ,Vascular disease ,business.industry ,Vascular biology ,Protein C Deficiency ,Exons ,Hematology ,Middle Aged ,medicine.disease ,Thrombosis ,Pedigree ,respiratory tract diseases ,Phenotype ,Codon, Nonsense ,Female ,business ,Protein C ,medicine.drug - Abstract
Hereditary basis of protein C deficiency (PCD) in thrombosis patients: First report from India
- Published
- 2009
41. Anti-HSP90 autoantibodies in sera of infertile women identify a dominant, conserved epitope EP6 (380-389) of HSP90 beta protein
- Author
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Asmita Choudhury, Eusebio S. Pires, Vrinda V. Khole, and Susan Idicula-Thomas
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lcsh:QH471-489 ,Peptide ,Autoantigens ,lcsh:Gynecology and obstetrics ,Epitope ,Endocrinology ,Antigen ,Heat shock protein ,Animals ,Humans ,lcsh:Reproduction ,HSP90 Heat-Shock Proteins ,lcsh:RG1-991 ,Autoantibodies ,chemistry.chemical_classification ,biology ,Immunodominant Epitopes ,Research ,Autoantibody ,Obstetrics and Gynecology ,Molecular biology ,Peptide Fragments ,Blot ,Reproductive Medicine ,chemistry ,Polyclonal antibodies ,biology.protein ,Female ,Rabbits ,Antibody ,Infertility, Female ,Oligopeptides ,Developmental Biology - Abstract
Background We earlier reported a simple specific test for detection of anti-ovarian antibodies in infertile women and identified number of specific molecular and cellular targets of which human heat shock protein 90-beta (HSP90 beta) was found to be the most immunodominant. The present study focuses on prediction and validation of the immunodominant epitope/s of this protein using sera from infertile women having anti-HSP90 autoantibodies. Methods Delineation of the immunodominant epitopes of HSP90 beta was done by using epitope prediction algorithms and 10 peptides (EP1-EP10) were custom synthesized. Their immunoreactivity was measured by ELISA using sera from patients and controls. To determine the most immunodominant epitope, the results were subjected to statistical analysis. The immunoreactivity of the immunodominant peptides were confirmed by dot blots using sera from patients. A rabbit polyclonal antibody against the immunodominant epitope was generated and its immunoreactivity to the parent protein in ovarian extracts as well in oocytes and embryos was investigated. Results Experimentally and statistically, peptide EP6 (380-389) seems to be the major antigenic epitope for the serum antibody binding followed by EP1 (1-12) and EP8 (488-498). Predicted 3D structures of these peptides demonstrated that they exist in the loop conformation which is the most mobile part of the protein. Also, analysis of the sequences of HSP90 beta across several species reveals that EP6 peptide forms a part of a well conserved motif. The polyclonal antibody generated to the immunodominant epitope- EP6 confirms similar biochemical and cellular immunoreactivity as seen with the patients' sera having anti-HSP90 autoantibodies. Conclusions The decapeptide EP6 is a major immunogenic epitope of HSP90 followed by EP1 and EP8. Knowledge of binding epitopes on the autoantigen is necessary to understand the subsequent pathologic events. The study might generate new tools for the detection of disease-inducing epitopes and a possible therapeutic intervention.
- Published
- 2011
42. ClotBase: a knowledgebase on proteins involved in blood coagulation
- Author
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Susan Idicula-Thomas, Ram Shankar Barai, Pravin Nilawe, and Archana Sonawani
- Subjects
Pathology ,medicine.medical_specialty ,Databases, Factual ,Protein Conformation ,Immunology ,Molecular Sequence Data ,Computational biology ,Biochemistry ,Consensus Sequence ,Databases, Genetic ,medicine ,Coagulation (water treatment) ,Animals ,Humans ,Amino Acid Sequence ,Databases, Protein ,Internet ,business.industry ,Cell Biology ,Hematology ,Blood Proteins ,Blood Coagulation Disorders ,Blood Coagulation Factors ,Disparate system ,Mutation ,business ,Sequence Alignment - Abstract
To the editor: Blood coagulation proteins (BCPs) play a major role in hemostasis.[1][1] Except for a few that have their own dedicated databases, information on most BCPs are scattered across various disparate data sources in multiple formats. This information has been compiled, manually curated
- Published
- 2010
43. Molecular basis of factor X deficiency cases from India
- Author
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Kanjaksha Ghosh, Leenam Mota, Susan Idicula-Thomas, and Shrimati Shetty
- Subjects
Adult ,Male ,Pediatrics ,medicine.medical_specialty ,Polymorphism, Genetic ,business.industry ,Amino Acids, Basic ,DNA Mutational Analysis ,India ,Hematology ,General Medicine ,Factor X deficiency ,Middle Aged ,Young Adult ,Phenotype ,Asian People ,Factor X ,Mutation ,Medicine ,Humans ,Female ,business ,Child ,Factor X Deficiency ,Genetics (clinical) - Published
- 2010
44. A novel 9-bp insertion detected in steroid 21-hydroxylase gene (CYP21A2): prediction of its structural and functional implications by computational methods
- Author
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Chinnaraj Saravanan, Sudhisha Dubey, R Raveendra Varma, Susan Idicula-Thomas, Anurupa Maitra, and Mohammad Anwaruddin
- Subjects
Models, Molecular ,congenital, hereditary, and neonatal diseases and abnormalities ,endocrine system diseases ,Steroid 21-Hydroxylase ,Protein Conformation ,medicine.medical_treatment ,Endocrinology, Diabetes and Metabolism ,Clinical Biochemistry ,DNA Mutational Analysis ,Molecular Sequence Data ,lcsh:Medicine ,Computational biology ,Biology ,urologic and male genital diseases ,Steroid ,Exon ,Protein structure ,medicine ,Humans ,Congenital adrenal hyperplasia ,Pharmacology (medical) ,Amino Acid Sequence ,Allele ,Peptide sequence ,Gene ,Molecular Biology ,Alleles ,Genetics ,Biochemistry, medical ,Adrenal Hyperplasia, Congenital ,Base Sequence ,Research ,Biochemistry (medical) ,lcsh:R ,nutritional and metabolic diseases ,Computational Biology ,Infant ,General Medicine ,Cell Biology ,Exons ,medicine.disease ,female genital diseases and pregnancy complications ,Mutagenesis, Insertional ,Female - Abstract
Background Steroid 21-hydroxylase deficiency is the most common cause of congenital adrenal hyperplasia (CAH). Detection of underlying mutations in CYP21A2 gene encoding steroid 21-hydroxylase enzyme is helpful both for confirmation of diagnosis and management of CAH patients. Here we report a novel 9-bp insertion in CYP21A2 gene and its structural and functional consequences on P450c21 protein by molecular modeling and molecular dynamics simulations methods. Methods A 30-day-old child was referred to our laboratory for molecular diagnosis of CAH. Sequencing of the entire CYP21A2 gene revealed a novel insertion (duplication) of 9-bp in exon 2 of one allele and a well-known mutation I172N in exon 4 of other allele. Molecular modeling and simulation studies were carried out to understand the plausible structural and functional implications caused by the novel mutation. Results Insertion of the nine bases in exon 2 resulted in addition of three valine residues at codon 71 of the P450c21 protein. Molecular dynamics simulations revealed that the mutant exhibits a faster unfolding kinetics and an overall destabilization of the structure due to the triple valine insertion was also observed. Conclusion The novel 9-bp insertion in exon 2 of CYP21A2 genesignificantly lowers the structural stability of P450c21 thereby leading to the probable loss of its function.
- Published
- 2008
45. Correlation between the structural stability and aggregation propensity of proteins
- Author
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Susan, Idicula-Thomas and Petety V, Balaji
- Subjects
Inclusion Bodies ,Amyloid ,Protein Folding ,Solubility ,Escherichia coli ,Proteins ,Protein Structure, Secondary ,Recombinant Proteins - Abstract
Protein aggregation, being an outcome of improper protein folding, is largely dependent on the folding kinetics of a protein. Previous studies have reported a positive correlation between the stability of the secondary structural elements of a protein and their rate of folding/unfolding. In this in silico study, the secondary and tertiary structures of proteins a) that form inclusion bodies on overexpression in Escherichia coli, b) that form amyloid fibrils and c) that are soluble on overexpression in E. coli are analyzed for certain features that are known to be associated with structural stability. The study revealed that the soluble proteins seem to have a higher rate of folding (based on contact order) and a lower percentage of exposed hydrophobic residues as compared to the inclusion body forming or amyloidogenic proteins. The soluble proteins also seem to have a more favored helix and strand composition (based on the known secondary structural propensities of amino acids). The secondary structure analyses also reveal that the evolutionary pressure is directed against protein aggregation. This understanding of the positive correlation between structural stability and solubility, along with the other parameters known to influence aggregation, could be exploited in the design of mutations aimed at reducing the aggregation propensity of the proteins.
- Published
- 2007
46. Identification of Novel Mutations in HEXA Gene in Children Affected with Tay Sachs Disease from India
- Author
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Swapnil Patil, Sarita Gupta, Susan Idicula-Thomas, Jayesh Sheth, Mehul Mistri, Frenny Sheth, Parag M Tamhankar, Pratima Kondurkar, and Daksha Sanghavi
- Subjects
Models, Molecular ,Protein Conformation ,Autosomal recessive ,Gene Identification and Analysis ,Developmental and Pediatric Neurology ,Ligands ,medicine.disease_cause ,Pediatrics ,Computational biology ,Consanguinity ,Human genetics ,Missense mutation ,Hexosaminidase ,Genetics ,Mutation ,Multidisciplinary ,Splice site mutation ,Tay-Sachs disease ,Neurodegenerative Diseases ,Founder Effect ,Neurology ,Child, Preschool ,Medicine ,Sequence Analysis ,Research Article ,Protein Binding ,Science ,Mutation, Missense ,India ,Genetic Counseling ,Biology ,Molecular Genetics ,Hexosaminidase A ,Genetic Mutation ,medicine ,Humans ,Family ,Clinical genetics ,Genetic Testing ,Base Sequence ,Point mutation ,Mutation Types ,DNA structure ,Infant ,Hydrogen Bonding ,HEXA ,medicine.disease ,Macromolecular structure analysis ,Haplotypes ,Protein structure ,Hexosaminidase activity - Abstract
Tay Sachs disease (TSD) is a neurodegenerative disorder due to β-hexosaminidase A deficiency caused by mutations in the HEXA gene. The mutations leading to Tay Sachs disease in India are yet unknown. We aimed to determine mutations leading to TSD in India by complete sequencing of the HEXA gene. The clinical inclusion criteria included neuroregression, seizures, exaggerated startle reflex, macrocephaly, cherry red spot on fundus examination and spasticity. Neuroimaging criteria included thalamic hyperdensities on CT scan/T1W images of MRI of the brain. Biochemical criteria included deficiency of hexosaminidase A (less than 2% of total hexosaminidase activity for infantile patients). Total leukocyte hexosaminidase activity was assayed by 4-methylumbelliferyl-N-acetyl-β-D-glucosamine lysis and hexosaminidase A activity was assayed by heat inactivation method and 4-methylumbelliferyl-N-acetyl-β-D-glucosamine-6-sulphate lysis method. The exons and exon-intron boundaries of the HEXA gene were bidirectionally sequenced using an automated sequencer. Mutations were confirmed in parents and looked up in public databases. In silico analysis for mutations was carried out using SIFT, Polyphen2, MutationT@ster and Accelrys Discovery Studio softwares. Fifteen families were included in the study. We identified six novel missense mutations, c.340 G>A (p.E114K), c.964 G>A (p.D322N), c.964 G>T (p.D322Y), c.1178C>G (p.R393P) and c.1385A>T (p.E462V), c.1432 G>A (p.G478R) and two previously reported mutations. c.1277_1278insTATC and c.508C>T (p.R170W). The mutation p.E462V was found in six unrelated families from Gujarat indicating a founder effect. A previously known splice site mutation c.805+1 G>C and another intronic mutation c.672+30 T>G of unknown significance were also identified. Mutations could not be identified in one family. We conclude that TSD patients from Gujarat should be screened for the common mutation p.E462V.
- Published
- 2012
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