177 results on '"Klein-Seetharaman J"'
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2. Solution NMR spectroscopy of [[[alpha]-.sup.15]N]lysine-labeled rhodopsin: the single peak observed in both conventional and TROSY-type HSQC spectra is ascribed to Lys-339 in the carboxyl-terminal peptide sequence
- Author
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Klein-Seetharaman, J., Reeves, P.J., Loewen, M.C., Getmanova, E.V., Chung, J., Schwalbe, H., Wright, P.E., and Khorana, H.G.
- Subjects
Rhodopsin -- Analysis ,Spectrum analysis -- Methods ,Science and technology - Abstract
[[[alpha-.sup.15]N]Lysine-labeled rhodopsin, prepared by expression of a synthetic gene in HEK293 cells, was investigated both by conventional and transverse relaxation optimized spectroscopy-type heteronuclear single quantum correlation spectroscopy. Whereas rhodopsin contains 11 lysines, 8 in cytoplasmic loops and 1 each in the C-terminal peptide sequence and the intradiscal and transmembrane domains, only a single sharp peak was observed in dodecyl maltoside micelles. This result did not change when dodecyl maltoside was replaced by octyl glucoside or octyl glucoside-phospholipid-mixed micelles. Additional signals of much lower and variable intensity appeared at temperatures above 20 [degrees] C and under denaturing conditions. Application of the transverse relaxation optimized spectroscopy sequence resulted in sharpening of resonances but also losses of signal intensity. The single peak observed has been assigned to the C-terminal Lys-339 from the following lines of evidence. First, the signal is observed in HNCO spectra of rhodopsin, containing the labeled [[sup.13]C]Ser-338/[[sup.15]N]Lys-339 dipeptide. Second, addition of a monoclonal anti-rhodopsin antibody that binds to the C-terminal 8 aa of rhodopsin caused disappearance of the peak. Third, truncated rhodopsin lacking the C-terminal sequence Asp-330-Ala-348 showed no signal, whereas the enzymatically produced peptide fragment containing the above sequence showed the single peak. The results indicate motion in the backbone amide groups of rhodopsin at time scales depending on their location in the sequence. At the C terminus, conformational averaging occurs at the nanosecond time scale but varies from microsecond to millisecond in other parts of the primary sequence. The motions reflecting conformational exchange may be general for membrane proteins containing transmembrane helical bundles.
- Published
- 2002
3. Early integration of the individual student in academic activities: A novel classroom concept for graduate education in molecular biophysics and structural biology
- Author
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Leuba, SH, Carney, SM, Dahlburg, EM, Eells, RJ, Ghodke, H, Yanamala, N, Schauer, G, Klein-Seetharaman, J, Leuba, SH, Carney, SM, Dahlburg, EM, Eells, RJ, Ghodke, H, Yanamala, N, Schauer, G, and Klein-Seetharaman, J
- Abstract
Background: A key challenge in interdisciplinary research is choosing the best approach from a large number of techniques derived from different disciplines and their interfaces. Results: To address this challenge in the area of Biophysics and Structural Biology, we have designed a graduate level course to teach students insightful use of experimental biophysical approaches in relationship to addressing biological questions related to biomolecular interactions and dynamics. A weekly seminar and data and literature club are used to compliment the training in class. The course contains wet-laboratory experimental demonstration and real-data analysis as well as lectures, grant proposal preparation and assessment, and student presentation components. Active student participation is mandatory in all aspects of the class. Students prepare materials for the class receiving individual and iterative feedback from course directors and local experts generating high quality classroom presentations. Conclusions: The ultimate goal of the course is to teach students the skills needed to weigh different experimental approaches against each other in addressing a specific biological question by thinking and executing academic tasks like faculty.
- Published
- 2014
4. Glutamic acid-rich proteins of rod photoreceptors are natively unfolded
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Batra-Safferling, R., Abarca-Heidemann, K., Körschen, H. G., Tziatzios, C., Stoldt, M., Budyak, I., Willbold, D., Schwalbe, H., Klein-Seetharaman, J., and Kaupp, U. B.
- Subjects
carbohydrates (lipids) ,ddc:570 - Abstract
Broadly neutralizing HIV antibodies (bNAbs) can recognize carbohydrate-dependent epitopes on gp120. In contrast to previously characterized glycan-dependent bNAbs that recognize high-mannose N-glycans, PGT121 binds complex-type N-glycans in glycan microarrays. We isolated the B-cell clone encoding PGT121, which segregates into PGT121-like and 10-1074-like groups distinguished by sequence, binding affinity, carbohydrate recognition, and neutralizing activity. Group 10-1074 exhibits remarkable potency and breadth but no detectable binding to protein-free glycans. Crystal structures of unliganded PGT121, 10-1074, and their likely germ-line precursor reveal that differential carbohydrate recognition maps to a cleft between complementarity determining region (CDR)H2 and CDRH3. This cleft was occupied by a complex-type N-glycan in a "liganded" PGT121 structure. Swapping glycan contact residues between PGT121 and 10-1074 confirmed their importance for neutralization. Although PGT121 binds complex-type N-glycans, PGT121 recognized high-mannose-only HIV envelopes in isolation and on virions. As HIV envelopes exhibit varying proportions of high-mannose- and complex-type N-glycans, these results suggest promiscuous carbohydrate interactions, an advantageous adaptation ensuring neutralization of all viruses within a given strain.
- Published
- 2006
5. Biodegradation of Single-Walled Carbon Nanotubes by Eosinophil Peroxidase
- Author
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Andõn, F. T., Kapralov, A. A., Yanamala, N., Feng, W., Baygan, Arjang, Chambers, B. J., Hultenby, K., Ye, Fei, Toprak, Muhammet S., Brandner, B. D., Fornara, Andrea, Klein-Seetharaman, J., Kotchey, G. P., Star, A., Shvedova, Anna A., Fadeel, B., Kagan, V. E., Andõn, F. T., Kapralov, A. A., Yanamala, N., Feng, W., Baygan, Arjang, Chambers, B. J., Hultenby, K., Ye, Fei, Toprak, Muhammet S., Brandner, B. D., Fornara, Andrea, Klein-Seetharaman, J., Kotchey, G. P., Star, A., Shvedova, Anna A., Fadeel, B., and Kagan, V. E.
- Abstract
Eosinophil peroxidase (EPO) is one of the major oxidant-producing enzymes during inflammatory states in the human lung. The degradation of single-walled carbon nanotubes (SWCNTs) upon incubation with human EPO and H2O 2 is reported. Biodegradation of SWCNTs is higher in the presence of NaBr, but neither EPO alone nor H2O2 alone caused the degradation of nanotubes. Molecular modeling reveals two binding sites for SWCNTs on EPO, one located at the proximal side (same side as the catalytic site) and the other on the distal side of EPO. The oxidized groups on SWCNTs in both cases are stabilized by electrostatic interactions with positively charged residues. Biodegradation of SWCNTs can also be executed in an ex vivo culture system using primary murine eosinophils stimulated to undergo degranulation. Biodegradation is proven by a range of methods including transmission electron microscopy, UV-visible-NIR spectroscopy, Raman spectroscopy, and confocal Raman imaging. Thus, human EPO (in vitro) and ex vivo activated eosinophils mediate biodegradation of SWCNTs: an observation that is relevant to pulmonary responses to these materials. Human eosinophil peroxidase (EPO) is able to degrade SWCNTs in vitro in the presence of H2O2. EPO is one of the major oxidant-generating enzymes present in human lungs during inflammatory states. The biodegradation of SWCNTs is evidenced also in an ex vivo culture system using primary murine eosinophils stimulated to undergo degranulation. These results are relevant to potential respiratory exposure to carbon nanotubes., QC 20131113
- Published
- 2013
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6. Comparative n-gram analysis of whole-genome sequences
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Ganapathiraju, Madhavi K, Weisser, D, Rosenfeld, R, Carbonell, J, Reddy, R, and Klein-Seetharaman, J
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Division of Biological Sciences ,BioInformatics Centre - Abstract
A current barrier for successful rational drug design is the lack of understanding of the structure space provided by the proteins in a cell that is determined by their sequence space. The protein sequences capable of folding to functional three-dimensional shapes of the proteins are clearly different for different organisms, since sequences obtained from human proteins often fail to form correct three-dimensional structures in bacterial organisms. In analogy to the question “What kind of things do people say?" we therefore need to ask the question “What kind of amino acid sequences occur in the proteins of an organism?" An understanding of the sequence space occupied by proteins in different organisms would have important applications for “translation" of proteins from the language of one organism into that of another and design of drugs that target sequences that might be unique or preferred by pathogenic organisms over those in human hosts. Here we describe the development of a biological language modeling toolkit (BLMT) for genome-wide statistical amino acid n-gram analysis and comparison across organisms (freely accessible at www.cs.cmu.edu/~blmt). Its functions were applied to 44 different bacterial, archaeal and the human genome. Amino acid n-gram distribution was found to be characteristic of organisms, as evidenced by (1) the ability of simple Markovian unigram models to distinguish organisms, (2) the marked variation in n-gram distributions across organisms above random variation, and (3) identification of organism-specific phrases in protein sequences that are greater than an order of magnitude standard deviations away from the mean. These lines of evidence suggest that different organisms utilize different “vocabularies" and “phrases", an observation that may provide novel approaches to drug development by specifically targeting these phrases. The results suggest that further detailed analysis of n-gram statistics of protein sequences from whole genomes will likely – in analogy to word n-gram analysis – result in powerful models for prediction, topic classification and information extraction of biological sequences.
- Published
- 2002
7. Human immunodeficiency virus type 1 Vpr: Oligomerization is an essential feature for its incorporation into virus particles
- Author
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Venkatachari, NJ, Walker, LA, Tastan, O, Le, T, Dempsey, TM, Li, Y, Yanamala, N, Srinivasan, A, Klein-Seetharaman, J, Montelaro, RC, Ayyavoo, V, Venkatachari, NJ, Walker, LA, Tastan, O, Le, T, Dempsey, TM, Li, Y, Yanamala, N, Srinivasan, A, Klein-Seetharaman, J, Montelaro, RC, and Ayyavoo, V
- Abstract
HIV-1 Vpr, a nonstructural viral protein associated with virus particles, has a positive role in the efficient transport of PIC into the nucleus of non-dividing target cells and enhances virus replication in primary T cells. Vpr is a 96 amino acid protein and the structure by NMR shows three helical domains. Vpr has been shown to exist as dimers and higher order oligomers. Considering the multifunctional nature of Vpr, the contribution of distinct helical domains to the dimer/oligomer structure of Vpr and the relevance of this feature to its functions are not clear. To address this, we have utilized molecular modeling approaches to identify putative models of oligomerization. The predicted interface residues were subjected to site-directed mutagenesis and evaluated their role in intermolecular interaction and virion incorporation. The interaction between Vpr molecules was monitored by Bimolecular Fluorescence complementation (BiFC) method. The results show that Vpr forms oligomers in live cells and residues in helical domains play critical roles in oligomerization. Interestingly, Vpr molecules defective in oligomerization also fail to incorporate into the virus particles. Based on the data, we suggest that oligomerization of Vpr is essential for virion incorporation property and may also have a role in the events associated with virus infection. © 2010 Venkatachari et al; licensee BioMed Central Ltd.
- Published
- 2010
8. Mitochondrial targeting of electron scavenging antioxidants: Regulation of selective oxidation vs random chain reactions
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Kagan, VE, Wipf, P, Stoyanovsky, D, Greenberger, JS, Borisenko, G, Belikova, NA, Yanamala, N, Samhan Arias, AK, Tungekar, MA, Jiang, J, Tyurina, YY, Ji, J, Klein-Seetharaman, J, Pitt, BR, Shvedova, AA, Bayir, H, Kagan, VE, Wipf, P, Stoyanovsky, D, Greenberger, JS, Borisenko, G, Belikova, NA, Yanamala, N, Samhan Arias, AK, Tungekar, MA, Jiang, J, Tyurina, YY, Ji, J, Klein-Seetharaman, J, Pitt, BR, Shvedova, AA, and Bayir, H
- Abstract
Effective regulation of highly compartmentalized production of reactive oxygen species and peroxidation reactions in mitochondria requires targeting of small molecule antioxidants and antioxidant enzymes into the organelles. This review describes recently developed approaches to mitochondrial targeting of small biologically active molecules based on: (i) preferential accumulation in mitochondria because of their hydrophobicity and positive charge (hydrophobic cations), (ii) binding with high affinity to an intra-mitochondrial constituent, and (iii) metabolic conversions by specific mitochondrial enzymes to reveal an active entity. In addition, targeted delivery of antioxidant enzymes via expression of leader sequences directing the proteins into mitochondria is considered. Examples of successful antioxidant and anti-apoptotic protection based on the ability of targeted cargoes to inhibit cytochrome c-catalyzed peroxidation of a mitochondria-specific phospholipid cardiolipin, in vitro and in vivo are presented. Particular emphasis is placed on the employment of triphenylphosphonium- and hemi-gramicidin S-moieties as two effective vehicles for mitochondrial delivery of antioxidants. © 2009 Elsevier B.V. All rights reserved.
- Published
- 2009
9. Mitochondrial Nm23-H4 can switch between phosphotransfer and lipid transfer activities
- Author
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Schlattner, U., primary, Amoscato, A., additional, Tyuina, Y.Y., additional, Tokarska-Schlattner, M., additional, Rios, S. Ramirez, additional, Boissan, M., additional, Epand, R.F., additional, Epand, R.M., additional, Klein-Seetharaman, J., additional, Lacombe, M.L., additional, and Kagan, V.E., additional
- Published
- 2012
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10. Binding of Ca2+ to Glutamic Acid-Rich Polypeptides from the Rod Outer Segment
- Author
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Haber-Pohlmeier, S., primary, Abarca-Heidemann, K., additional, Körschen, H.G., additional, Dhiman, H. Kaur, additional, Heberle, J., additional, Schwalbe, H., additional, Klein-Seetharaman, J., additional, Kaupp, U.B., additional, and Pohlmeier, A., additional
- Published
- 2007
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11. Characterization of protein secondary structure
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Ganapathiraju, M.K., primary, Klein-Seetharaman, J., additional, Balakrishnan, N., additional, and Reddy, R., additional
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- 2004
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12. Analogies between structural and systems biology and systems-of-systems engineering in dynamic environments.
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Moitra, S., Yanamala, N., Tastan, O., Singh, I., Langmead, C.J., and Klein-Seetharaman, J.
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- 2010
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13. Communication interception of human signal transduction pathways by Human Immunodeficiency Virus-1.
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Balakrishnan, S., Tastan, O., Carbonell, J.G., and Klein-Seetharaman, J.
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- 2009
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14. Comparative n-gram analysis of whole-genome protein sequences
- Author
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Ganapathiraju, M., primary, Weisser, D., additional, Rosenfeld, R., additional, Carbonell, J., additional, Reddy, R., additional, and Klein-Seetharaman, J., additional
- Published
- 2002
- Full Text
- View/download PDF
15. Solution NMR spectroscopy of [α−[sup 15]N]lysine-labeled rhodopsin: The single peak observed in both conventional and TROSY-type HSQC spectra is ascribed to Lys-339 in th carboxyl-terminal reptide sequence.
- Author
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Klein-Seetharaman, J., Reeves, P.J., Loewen, M.C., Getmanova, E.V., Chung, J., Schwalbe, H., and Wright, P.E.
- Subjects
- *
RHODOPSIN , *GLUCOSIDES , *NUCLEAR magnetic resonance - Abstract
Examines the [α15N] Lysine-labeled rhodopsin by conventional and transverse relaxation optimized spectroscopy-type heteronuclear single quantum correlation spectroscopy. Replacement of dodecyl maltoside by octyl glucoside; Use of nuclear magnetic resonance spectroscopy; Preparation of HEK1935 cell lines with opsin genes.
- Published
- 2002
- Full Text
- View/download PDF
16. Flexibility of the cytoplasmic domain of the phototaxis transducer II from Natronomonas pharaonis
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Il, Budyak, Os, Mironova, Yanamala N, Manoharan V, Büldt G, Ramona Schlesinger, and Klein-Seetharaman J
17. Alternative paths in HIV-1 targeted human signal transduction pathways
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Klein-Seetharaman Judith, Carbonell Jaime, Tastan Oznur, and Balakrishnan Sivaraman
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Biotechnology ,TP248.13-248.65 ,Genetics ,QH426-470 - Abstract
Abstract Background Human immunodeficiency virus-1 (HIV-1) has a minimal genome of only 9 genes, which encode 15 proteins. HIV-1 thus depends on the human host for virtually every aspect of its life cycle. The universal language of communication in biological systems, including between pathogen and host, is via signal transduction pathways. The fundamental units of these pathways are protein protein interactions. Understanding the functional significance of HIV-1, human interactions requires viewing them in the context of human signal transduction pathways. Results Integration of HIV-1, human interactions with known signal transduction pathways indicates that the majority of known human pathways have the potential to be effected through at least one interaction with an HIV-1 protein at some point during the HIV-1 life cycle. For each pathway, we define simple paths between start points (i.e. no edges going into a node) and end points (i.e. no edges leaving a node). We then identify the paths that pass through human proteins that interact with HIV-1 proteins. We supplement the combined map with functional information, including which proteins are known drug targets and which proteins contribute significantly to HIV-1 function as revealed by recent siRNA screens. We find that there are often alternative paths starting and ending at the same proteins but circumventing the intermediate steps disrupted by HIV-1. Conclusion A mapping of HIV-1, human interactions to human signal transduction pathways is presented here to link interactions with functions. We proposed a new way of analyzing the virus host interactions by identifying HIV-1 targets as well as alternative paths bypassing the HIV-1 targeted steps. This approach yields numerous experimentally testable hypotheses on how HIV-1 function may be compromised and human cellular function restored by pharmacological approaches. We are making the full set of pathway analysis results available to the community.
- Published
- 2009
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18. Preferential binding of allosteric modulators to active and inactive conformational states of metabotropic glutamate receptors
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Klein-Seetharaman Judith, Tirupula Kalyan C, and Yanamala Naveena
- Subjects
Computer applications to medicine. Medical informatics ,R858-859.7 ,Biology (General) ,QH301-705.5 - Abstract
Abstract Metabotropic glutamate receptors (mGluRs) are G protein coupled receptors that play important roles in synaptic plasticity and other neuro-physiological and pathological processes. Allosteric mGluR ligands are particularly promising drug targets because of their modulatory effects – enhancing or suppressing the response of mGluRs to glutamate. The mechanism by which this modulation occurs is not known. Here, we propose the hypothesis that positive and negative modulators will differentially stabilize the active and inactive conformations of the receptors, respectively. To test this hypothesis, we have generated computational models of the transmembrane regions of different mGluR subtypes in two different conformations. The inactive conformation was modeled using the crystal structure of the inactive, dark state of rhodopsin as template and the active conformation was created based on a recent model of the light-activated state of rhodopsin. Ligands for which the nature of their allosteric effects on mGluRs is experimentally known were docked to the modeled mGluR structures using ArgusLab and Autodock softwares. We find that the allosteric ligand binding pockets of mGluRs are overlapping with the retinal binding pocket of rhodopsin, and that ligands have strong preferences for the active and inactive states depending on their modulatory nature. In 8 out of 14 cases (57%), the negative modulators bound the inactive conformations with significant preference using both docking programs, and 6 out of 9 cases (67%), the positive modulators bound the active conformations. Considering results by the individual programs only, even higher correlations were observed: 12/14 (86%) and 8/9 (89%) for ArgusLab and 10/14 (71%) and 7/9 (78%) for AutoDock. These findings strongly support the hypothesis that mGluR allosteric modulation occurs via stabilization of different conformations analogous to those identified in rhodopsin where they are induced by photochemical isomerization of the retinal ligand – despite the extensive differences in sequences between mGluRs and rhodopsin.
- Published
- 2008
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19. Human immunodeficiency virus type 1 Vpr: oligomerization is an essential feature for its incorporation into virus particles
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Klein-Seetharaman Judith, Srinivasan Alagarsamy, Yanamala Naveena, Li Yaming, Dempsey Timothy M, Le Thien, Tastan Oznur, Walker Leah A, Venkatachari Narasimhan J, Montelaro Ronald C, and Ayyavoo Velpandi
- Subjects
Infectious and parasitic diseases ,RC109-216 - Abstract
Abstract HIV-1 Vpr, a nonstructural viral protein associated with virus particles, has a positive role in the efficient transport of PIC into the nucleus of non-dividing target cells and enhances virus replication in primary T cells. Vpr is a 96 amino acid protein and the structure by NMR shows three helical domains. Vpr has been shown to exist as dimers and higher order oligomers. Considering the multifunctional nature of Vpr, the contribution of distinct helical domains to the dimer/oligomer structure of Vpr and the relevance of this feature to its functions are not clear. To address this, we have utilized molecular modeling approaches to identify putative models of oligomerization. The predicted interface residues were subjected to site-directed mutagenesis and evaluated their role in intermolecular interaction and virion incorporation. The interaction between Vpr molecules was monitored by Bimolecular Fluorescence complementation (BiFC) method. The results show that Vpr forms oligomers in live cells and residues in helical domains play critical roles in oligomerization. Interestingly, Vpr molecules defective in oligomerization also fail to incorporate into the virus particles. Based on the data, we suggest that oligomerization of Vpr is essential for virion incorporation property and may also have a role in the events associated with virus infection.
- Published
- 2010
- Full Text
- View/download PDF
20. Navigating Challenges and Opportunities in Multi-Omics Integration for Personalized Healthcare.
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Mohr AE, Ortega-Santos CP, Whisner CM, Klein-Seetharaman J, and Jasbi P
- Abstract
The field of multi-omics has witnessed unprecedented growth, converging multiple scientific disciplines and technological advances. This surge is evidenced by a more than doubling in multi-omics scientific publications within just two years (2022-2023) since its first referenced mention in 2002, as indexed by the National Library of Medicine. This emerging field has demonstrated its capability to provide comprehensive insights into complex biological systems, representing a transformative force in health diagnostics and therapeutic strategies. However, several challenges are evident when merging varied omics data sets and methodologies, interpreting vast data dimensions, streamlining longitudinal sampling and analysis, and addressing the ethical implications of managing sensitive health information. This review evaluates these challenges while spotlighting pivotal milestones: the development of targeted sampling methods, the use of artificial intelligence in formulating health indices, the integration of sophisticated n -of-1 statistical models such as digital twins, and the incorporation of blockchain technology for heightened data security. For multi-omics to truly revolutionize healthcare, it demands rigorous validation, tangible real-world applications, and smooth integration into existing healthcare infrastructures. It is imperative to address ethical dilemmas, paving the way for the realization of a future steered by omics-informed personalized medicine.
- Published
- 2024
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21. PRDX6 contributes to selenocysteine metabolism and ferroptosis resistance.
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Chen Z, Inague A, Kaushal K, Fazeli G, N Xavier da Silva T, Ferreira Dos Santos A, Cheytan T, Porto Freitas F, Yildiz U, Gasparello Viviani L, Santiago Lima R, Peglow Pinz M, Medeiros I, Geronimo Pires Alegria T, Pereira da Silva R, Regina Diniz L, Weinzweig S, Klein-Seetharaman J, Trumpp A, Manas A, Hondal R, Fischer M, Bartenhagen C, Shimada BK, Seale LA, Fabiano M, Schweizer U, Netto LE, Meotti FC, Alborzinia H, Miyamoto S, and Friedmann Angeli JP
- Abstract
Selenocysteine (Sec) metabolism is crucial for cellular function and ferroptosis prevention and has traditionally been thought to begin with the uptake of the Sec carrier selenoprotein P (SELENOP). Following uptake, Sec released from SELENOP undergoes metabolisation via selenocysteine lyase (SCLY), producing selenide, a substrate used by selenophosphate synthetase 2 (SEPHS2), which provides the essential selenium donor - selenophosphate - for the biosynthesis of the selenocysteine tRNA. Here, we report the discovery of an alternative pathway mediating Sec metabolisation that is independent of SCLY and mediated by peroxiredoxin 6 (PRDX6). Mechanistically, we demonstrate that PRDX6 can readily react with selenide and interact with SEPHS2, potentially acting as a selenium delivery system. Moreover, we demonstrate the presence and functional significance of this alternative route in cancer cells where we reveal a notable association between elevated expression of PRDX6 with a highly aggressive neuroblastoma subtype. Altogether, our study sheds light on a previously unrecognized aspect of Sec metabolism and its implications in ferroptosis, offering new avenues for therapeutic exploitation.
- Published
- 2024
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22. Catalytic role of histidine-114 in the hydrolytic dehalogenation of chlorothalonil by Pseudomonas sp. CTN-3.
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Gerlich G, Miller C, Yang X, Diviesti K, Bennett B, Klein-Seetharaman J, and Holz RC
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- Hydrolysis, Biocatalysis, Catalytic Domain, Fungicides, Industrial chemistry, Fungicides, Industrial metabolism, Halogenation, Hydrolases metabolism, Hydrolases chemistry, Pseudomonas enzymology, Pseudomonas metabolism, Nitriles metabolism, Nitriles chemistry, Histidine chemistry, Histidine metabolism
- Abstract
Chlorothalonil (2,4,5,6-tetrachloroisophthalonitrile; TPN) is an environmentally persistent fungicide that sees heavy use in the USA and is highly toxic to aquatic species and birds, as well as a probable human carcinogen. The chlorothalonil dehalogenase from Pseudomonas sp. CTN-3 (Chd, UniProtKB C9EBR5) degrades TPN to its less toxic 4-OH-TPN analog making it an exciting candidate for the development of a bioremediation process for TPN; however, little is currently known about its catalytic mechanism. Therefore, an active site residue histidine-114 (His114) which forms a hydrogen bond with the Zn(II)-bound water/hydroxide and has been suggested to be the active site acid/base, was substituted by an Ala residue. Surprisingly, Chd
H114A exhibited catalytic activity with a kcat value of 1.07 s-1 , ~ 5% of wild-type (WT) Chd, and a KM of 32 µM. Thus, His114 is catalytically important but not essential. The electronic and structural aspects of the WT Chd and ChdH114A active sites were examined using UV-Vis and EPR spectroscopy on the catalytically competent Co(II)-substituted enzyme as well as all-atomistic molecular dynamics (MD) simulations. Combination of these data suggest His114 can quickly and reversibly move nearly 2 Å between one conformation that facilitates catalysis and another that enables product egress and active site recharge. In light of experimental and computational data on ChdH114A , Asn216 appears to play a role in substrate binding and preorganization of the transition-state while Asp116 likely facilitates the deprotonation of the Zn(II)-bound water in the absence of His114. Based on these data, an updated proposed catalytic mechanism for Chd is presented., (© 2024. The Author(s), under exclusive licence to Society for Biological Inorganic Chemistry (SBIC).)- Published
- 2024
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23. Gut microbiome remodeling and metabolomic profile improves in response to protein pacing with intermittent fasting versus continuous caloric restriction.
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Mohr AE, Sweazea KL, Bowes DA, Jasbi P, Whisner CM, Sears DD, Krajmalnik-Brown R, Jin Y, Gu H, Klein-Seetharaman J, Arciero KM, Gumpricht E, and Arciero PJ
- Subjects
- Humans, Male, Female, Adult, Middle Aged, Feces microbiology, Feces chemistry, Metabolome, Weight Loss physiology, Obesity metabolism, Obesity therapy, Obesity diet therapy, Obesity microbiology, Dietary Proteins metabolism, Dietary Proteins administration & dosage, Intermittent Fasting, Gastrointestinal Microbiome physiology, Caloric Restriction methods, Fasting blood, Body Composition, Metabolomics methods
- Abstract
The gut microbiome (GM) modulates body weight/composition and gastrointestinal functioning; therefore, approaches targeting resident gut microbes have attracted considerable interest. Intermittent fasting (IF) and protein pacing (P) regimens are effective in facilitating weight loss (WL) and enhancing body composition. However, the interrelationships between IF- and P-induced WL and the GM are unknown. The current randomized controlled study describes distinct fecal microbial and plasma metabolomic signatures between combined IF-P (n = 21) versus a heart-healthy, calorie-restricted (CR, n = 20) diet matched for overall energy intake in free-living human participants (women = 27; men = 14) with overweight/obesity for 8 weeks. Gut symptomatology improves and abundance of Christensenellaceae microbes and circulating cytokines and amino acid metabolites favoring fat oxidation increase with IF-P (p < 0.05), whereas metabolites associated with a longevity-related metabolic pathway increase with CR (p < 0.05). Differences indicate GM and metabolomic factors play a role in WL maintenance and body composition. This novel work provides insight into the GM and metabolomic profile of participants following an IF-P or CR diet and highlights important differences in microbial assembly associated with WL and body composition responsiveness. These data may inform future GM-focused precision nutrition recommendations using larger sample sizes of longer duration. Trial registration, March 6, 2020 (ClinicalTrials.gov as NCT04327141), based on a previous randomized intervention trial., (© 2024. The Author(s).)
- Published
- 2024
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24. 7-Dehydrocholesterol is an endogenous suppressor of ferroptosis.
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Freitas FP, Alborzinia H, Dos Santos AF, Nepachalovich P, Pedrera L, Zilka O, Inague A, Klein C, Aroua N, Kaushal K, Kast B, Lorenz SM, Kunz V, Nehring H, Xavier da Silva TN, Chen Z, Atici S, Doll SG, Schaefer EL, Ekpo I, Schmitz W, Horling A, Imming P, Miyamoto S, Wehman AM, Genaro-Mattos TC, Mirnics K, Kumar L, Klein-Seetharaman J, Meierjohann S, Weigand I, Kroiss M, Bornkamm GW, Gomes F, Netto LES, Sathian MB, Konrad DB, Covey DF, Michalke B, Bommert K, Bargou RC, Garcia-Saez A, Pratt DA, Fedorova M, Trumpp A, Conrad M, and Friedmann Angeli JP
- Subjects
- Animals, Humans, Cell Survival, Lipid Peroxidation, Neoplasm Transplantation, Oxidation-Reduction, Phenotype, Reproducibility of Results, Burkitt Lymphoma metabolism, Burkitt Lymphoma pathology, Dehydrocholesterols metabolism, Ferroptosis, Neuroblastoma metabolism, Neuroblastoma pathology
- Abstract
Ferroptosis is a form of cell death that has received considerable attention not only as a means to eradicate defined tumour entities but also because it provides unforeseen insights into the metabolic adaptation that tumours exploit to counteract phospholipid oxidation
1,2 . Here, we identify proferroptotic activity of 7-dehydrocholesterol reductase (DHCR7) and an unexpected prosurvival function of its substrate, 7-dehydrocholesterol (7-DHC). Although previous studies suggested that high concentrations of 7-DHC are cytotoxic to developing neurons by favouring lipid peroxidation3 , we now show that 7-DHC accumulation confers a robust prosurvival function in cancer cells. Because of its far superior reactivity towards peroxyl radicals, 7-DHC effectively shields (phospho)lipids from autoxidation and subsequent fragmentation. We provide validation in neuroblastoma and Burkitt's lymphoma xenografts where we demonstrate that the accumulation of 7-DHC is capable of inducing a shift towards a ferroptosis-resistant state in these tumours ultimately resulting in a more aggressive phenotype. Conclusively, our findings provide compelling evidence of a yet-unrecognized antiferroptotic activity of 7-DHC as a cell-intrinsic mechanism that could be exploited by cancer cells to escape ferroptosis., (© 2024. The Author(s), under exclusive licence to Springer Nature Limited.)- Published
- 2024
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25. Insulin signaling and pharmacology in humans and in corals.
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Murthy MHS, Jasbi P, Lowe W, Kumar L, Olaosebikan M, Roger L, Yang J, Lewinski N, Daniels N, Cowen L, and Klein-Seetharaman J
- Subjects
- Animals, Humans, Insulin metabolism, Pancreas metabolism, Signal Transduction, Anthozoa metabolism, Islets of Langerhans metabolism
- Abstract
Once thought to be a unique capability of the Langerhans islets in the pancreas of mammals, insulin (INS) signaling is now recognized as an evolutionarily ancient function going back to prokaryotes. INS is ubiquitously present not only in humans but also in unicellular eukaryotes, fungi, worms, and Drosophila . Remote homologue identification also supports the presence of INS and INS receptor in corals where the availability of glucose is largely dependent on the photosynthetic activity of the symbiotic algae. The cnidarian animal host of corals operates together with a 20,000-sized microbiome, in direct analogy to the human gut microbiome. In humans, aberrant INS signaling is the hallmark of metabolic disease, and is thought to play a major role in aging, and age-related diseases, such as Alzheimer's disease. We here would like to argue that a broader view of INS beyond its human homeostasis function may help us understand other organisms, and in turn, studying those non-model organisms may enable a novel view of the human INS signaling system. To this end, we here review INS signaling from a new angle, by drawing analogies between humans and corals at the molecular level., Competing Interests: The authors declare that they have no competing interests., (© 2024 Murthy et al.)
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- 2024
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26. Creation of a Curated Database of Experimentally Determined Human Protein Structures for the Identification of Its Targetome.
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Ovanessians A, Snow C, Jennewein T, Sarkar S, Speyer G, and Klein-Seetharaman J
- Subjects
- Humans, Ligands, Binding Sites, Protein Binding, Databases, Protein, Computational Biology, Proteins chemistry
- Abstract
Assembling an "integrated structural map of the human cell" at atomic resolution will require a complete set of all human protein structures available for interaction with other biomolecules - the human protein structure targetome - and a pipeline of automated tools that allow quantitative analysis of millions of protein-ligand interactions. Toward this goal, we here describe the creation of a curated database of experimentally determined human protein structures. Starting with the sequences of 20,422 human proteins, we selected the most representative structure for each protein (if available) from the protein database (PDB), ranking structures by coverage of sequence by structure, depth (the difference between the final and initial residue number of each chain), resolution, and experimental method used to determine the structure. To enable expansion into an entire human targetome, we docked small molecule ligands to our curated set of protein structures. Using design constraints derived from comparing structure assembly and ligand docking results obtained with challenging protein examples, we here propose to combine this curated database of experimental structures with AlphaFold predictions and multi-domain assembly using DEMO2 in the future. To demonstrate the utility of our curated database in identification of the human protein structure targetome, we used docking with AutoDock Vina and created tools for automated analysis of affinity and binding site locations of the thousands of protein-ligand prediction results. The resulting human targetome, which can be updated and expanded with an evolving curated database and increasing numbers of ligands, is a valuable addition to the growing toolkit of structural bioinformatics.
- Published
- 2024
27. Inflammation and the pathological progression of Alzheimer's disease are associated with low circulating choline levels.
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Judd JM, Jasbi P, Winslow W, Serrano GE, Beach TG, Klein-Seetharaman J, and Velazquez R
- Subjects
- Humans, Mice, Animals, Choline pharmacology, Tumor Necrosis Factor-alpha, Plaque, Amyloid pathology, Amyloid beta-Peptides metabolism, Acetylcholine, Inflammation, tau Proteins metabolism, Alzheimer Disease pathology
- Abstract
Deficiency of dietary choline, an essential nutrient, is observed worldwide, with ~ 90% of Americans being deficient. Previous work highlights a relationship between decreased choline intake and an increased risk for cognitive decline and Alzheimer's disease (AD). The associations between blood circulating choline and the pathological progression in both mild cognitive impairment (MCI) and AD remain unknown. Here, we examined these associations in a cohort of patients with MCI with presence of either sparse or high neuritic plaque density and Braak stage and a second cohort with either moderate AD (moderate to frequent neuritic plaques, Braak stage = IV) or severe AD (frequent neuritic plaques, Braak stage = VI), compared to age-matched controls. Metabolomic analysis was performed on serum from the AD cohort. We then assessed the effects of dietary choline deficiency (Ch-) in 3xTg-AD mice and choline supplementation (Ch+) in APP/PS1 mice, two rodent models of AD. The levels of circulating choline were reduced while pro-inflammatory cytokine TNFα was elevated in serum of both MCI sparse and high pathology cases. Reduced choline and elevated TNFα correlated with higher neuritic plaque density and Braak stage. In AD patients, we found reductions in choline, its derivative acetylcholine (ACh), and elevated TNFα. Choline and ACh levels were negatively correlated with neuritic plaque load, Braak stage, and TNFα, but positively correlated with MMSE, and brain weight. Metabolites L-Valine, 4-Hydroxyphenylpyruvic, Methylmalonic, and Ferulic acids were significantly associated with circuiting choline levels. In 3xTg-AD mice, the Ch- diet increased amyloid-β levels and tau phosphorylation in cortical tissue, and TNFα in both blood and cortical tissue, paralleling the severe human-AD profile. Conversely, the Ch+ diet increased choline and ACh while reducing amyloid-β and TNFα levels in brains of APP/PS1 mice. Collectively, low circulating choline is associated with AD-neuropathological progression, illustrating the importance of adequate dietary choline intake to offset disease., (© 2023. The Author(s).)
- Published
- 2023
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28. Precision Medicine as a New Frontier in Speech-Language Pathology: How Applying Insights From Behavior Genomics Can Improve Outcomes in Communication Disorders.
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Peter B, Bruce L, Finestack L, Dinu V, Wilson M, Klein-Seetharaman J, Lewis CR, Braden BB, Tang YY, Scherer N, VanDam M, and Potter N
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- Humans, Speech, Precision Medicine, Genomics, Communication, Communication Disorders genetics, Speech-Language Pathology education
- Abstract
Purpose: Precision medicine is an emerging intervention paradigm that leverages knowledge of risk factors such as genotypes, lifestyle, and environment toward proactive and personalized interventions. Regarding genetic risk factors, examples of interventions informed by the field of medical genomics are pharmacological interventions tailored to an individual's genotype and anticipatory guidance for children whose hearing impairment is predicted to be progressive. Here, we show how principles of precision medicine and insights from behavior genomics have relevance for novel management strategies of behaviorally expressed disorders, especially disorders of spoken language., Method: This tutorial presents an overview of precision medicine, medical genomics, and behavior genomics; case examples of improved outcomes; and strategic goals toward enhancing clinical practice., Results: Speech-language pathologists (SLPs) see individuals with various communication disorders due to genetic variants. Ways of using insights from behavior genomics and implementing principles of precision medicine include recognizing early signs of undiagnosed genetic disorders in an individual's communication patterns, making appropriate referrals to genetics professionals, and incorporating genetic findings into management plans. Patients benefit from a genetics diagnosis by gaining a deeper and more prognostic understanding of their condition, obtaining more precisely targeted interventions, and learning about their recurrence risks., Conclusions: SLPs can achieve improved outcomes by expanding their purview to include genetics. To drive this new interdisciplinary framework forward, goals should include systematic training in clinical genetics for SLPs, enhanced understanding of genotype-phenotype associations, leveraging insights from animal models, optimizing interprofessional team efforts, and developing novel proactive and personalized interventions.
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- 2023
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29. Low circulating choline, a modifiable dietary factor, is associated with the pathological progression and metabolome dysfunction in Alzheimer's disease.
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Judd JM, Jasbi P, Winslow W, Serrano GE, Beach TG, Klein-Seetharaman J, and Velazquez R
- Abstract
Most Americans (∼90%) are deficient in dietary choline, an essential nutrient. Associations between circulating choline and pathological progression in Alzheimer's disease (AD) remain unknown. Here, we examined these associations and performed a metabolomic analysis in blood serum from severe AD, moderate AD, and healthy controls. Additionally, to gain mechanistic insight, we assessed the effects of dietary choline deficiency (Ch-) in 3xTg-AD mice and choline supplementation (Ch+) in APP/PS1 mice. In humans, we found AD-associated reductions in choline, it's derivative acetylcholine (ACh), and elevated pro-inflammatory cytokine TNFα. Choline and ACh were negatively correlated with Plaque density, Braak stage, and TNFα, but positively correlated with MMSE and brain weight. Metabolites L-Valine, 4-Hydroxyphenylpyruvic, Methylmalonic, and Ferulic acids were associated with choline levels. In mice, Ch-paralleled AD severe, but Ch+ was protective. In conclusion, low circulating choline is associated with AD-neuropathological progression, illustrating the importance of dietary choline consumption to offset disease.
- Published
- 2023
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30. High-frequency imagery to capture coral tissue (Montipora capricornis) response to environmental stress, a pilot study.
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Li S, Roger LM, Kumar L, Lewinski NA, Klein-Seetharaman J, Putnam HM, and Yang J
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- Animals, Pilot Projects, Coral Reefs, Stress, Physiological, Symbiosis physiology, Anthozoa physiology
- Abstract
Environment stress is a major threat to the existence of coral reefs and has generated a lot of interest in the coral research community. Under the environmental stress, corals can experience tissue loss and/or the breakdown of symbiosis between the cnidarian host and its symbiotic algae causing the coral tissue to appear white as the skeleton can be seen by transparency. Image analysis is a common method used to assess tissue response under the environmental stress. However, the traditional approach is limited by the dynamic nature of the coral-algae symbiosis. Here, we observed coral tissue response in the scleractinian coral, Montipora capricornis, using high frequency image analysis throughout the experiment, as opposed to the typical start/end point assessment method. Color analysis reveals that the process can be divided into five stages with two critical stages according to coral tissue morphology and color ratio. We further explore changes to the morphology of individual polyps by means of the Pearson correlation coefficient and recurrence plots, where the quasi-periodic and nonstationary dynamics can be identified. The recurrence quantification analysis also allows the comparison between the different polyps. Our research provides a detailed visual and mathematical analysis of coral tissue response to environmental stress, which potentially shows universal applicability. Moreover, our approach provides a robust quantitative advancement for improving our insight into a suite of biotic responses in the perspective of coral health evaluation and fate prediction., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2023 Li et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)
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- 2023
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31. Gobind's last graduate student.
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Klein-Seetharaman J
- Abstract
Written on the occasion of his 100th birthday, this is a personal account of my time as a graduate student with Nobel laureate, H. Gobind Khorana, at the Massachusetts Institute of Technology from 1996 to 2000., Competing Interests: Conflict of interestNot applicable., (© International Union for Pure and Applied Biophysics (IUPAB) and Springer-Verlag GmbH Germany, part of Springer Nature 2023.)
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- 2023
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32. Transfer of knowledge from model organisms to evolutionarily distant non-model organisms: The coral Pocillopora damicornis membrane signaling receptome.
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Kumar L, Brenner N, Sledzieski S, Olaosebikan M, Roger LM, Lynn-Goin M, Klein-Seetharaman R, Berger B, Putnam H, Yang J, Lewinski NA, Singh R, Daniels NM, Cowen L, and Klein-Seetharaman J
- Subjects
- Animals, Humans, Genome, Models, Animal, Anthozoa genetics, Anthozoa physiology, Receptors, G-Protein-Coupled genetics, Receptors, G-Protein-Coupled metabolism, Signal Transduction
- Abstract
With the ease of gene sequencing and the technology available to study and manipulate non-model organisms, the extension of the methodological toolbox required to translate our understanding of model organisms to non-model organisms has become an urgent problem. For example, mining of large coral and their symbiont sequence data is a challenge, but also provides an opportunity for understanding functionality and evolution of these and other non-model organisms. Much more information than for any other eukaryotic species is available for humans, especially related to signal transduction and diseases. However, the coral cnidarian host and human have diverged over 700 million years ago and homologies between proteins in the two species are therefore often in the gray zone, or at least often undetectable with traditional BLAST searches. We introduce a two-stage approach to identifying putative coral homologues of human proteins. First, through remote homology detection using Hidden Markov Models, we identify candidate human homologues in the cnidarian genome. However, for many proteins, the human genome alone contains multiple family members with similar or even more divergence in sequence. In the second stage, therefore, we filter the remote homology results based on the functional and structural plausibility of each coral candidate, shortlisting the coral proteins likely to have conserved some of the functions of the human proteins. We demonstrate our approach with a pipeline for mapping membrane receptors in humans to membrane receptors in corals, with specific focus on the stony coral, P. damicornis. More than 1000 human membrane receptors mapped to 335 coral receptors, including 151 G protein coupled receptors (GPCRs). To validate specific sub-families, we chose opsin proteins, representative GPCRs that confer light sensitivity, and Toll-like receptors, representative non-GPCRs, which function in the immune response, and their ability to communicate with microorganisms. Through detailed structure-function analysis of their ligand-binding pockets and downstream signaling cascades, we selected those candidate remote homologues likely to carry out related functions in the corals. This pipeline may prove generally useful for other non-model organisms, such as to support the growing field of synthetic biology., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2023 Kumar et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)
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- 2023
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33. Evolution of gas in scattering media absorption spectroscopy as a neonatal pulmonary monitoring device.
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Muniraman HK, Klein-Seetharaman J, and Bhandari V
- Subjects
- Humans, Infant, Newborn, Spectrum Analysis methods, Scattering, Radiation, Lung
- Published
- 2022
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34. Evaluation of fluorescence-based viability stains in cells dissociated from scleractinian coral Pocillopora damicornis.
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Roger LM, Adarkwa Darko Y, Bernas T, White F, Olaosebikan M, Cowen L, Klein-Seetharaman J, and Lewinski NA
- Subjects
- Animals, Chlorophyll metabolism, Fluorescent Dyes metabolism, Mammals, Staining and Labeling, Anthozoa metabolism, Insulins metabolism
- Abstract
The application of established cell viability assays such as the commonly used trypan blue staining method to coral cells is not straightforward due to different culture parameters and different cellular features specific to mammalian cells compared to marine invertebrates. Using Pocillopora damicornis as a model, we characterized the autofluorescence and tested different fluorescent dye pair combinations to identify alternative viability indicators. The cytotoxicity of different representative molecules, namely small organic molecules, proteins and nanoparticles (NP), was measured after 24 h of exposure using the fluorescent dye pair Hoechst 33342 and SYTOX orange. Our results show that this dye pair can be distinctly measured in the presence of fluorescent proteins plus chlorophyll. P. damicornis cells exposed for 24 h to Triton-X100, insulin or titanium dioxide (TiO
2 ) NPs, respectively, at concentrations ranging from 0.5 to 100 µg/mL, revealed a LC50 of 0.46 µg/mL for Triton-X100, 6.21 µg/mL for TiO2 NPs and 33.9 µg/mL for insulin. This work presents the approach used to customize dye pairs for membrane integrity-based cell viability assays considering the species- and genotype-specific autofluorescence of scleractinian corals, namely: endogenous fluorescence characterization followed by the selection of dyes that do not overlap with endogenous signals., (© 2022. The Author(s).)- Published
- 2022
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35. Structure-based survey of ligand binding in the human insulin receptor.
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Kumar L, Vizgaudis W, and Klein-Seetharaman J
- Subjects
- Cryoelectron Microscopy, Humans, Ligands, Protein Binding, Insulin, Receptor, Insulin
- Abstract
The insulin receptor is a membrane protein responsible for the regulation of nutrient balance; and therefore, it is an attractive target in the treatment of diabetes and metabolic syndrome. Pharmacology of the insulin receptor involves two distinct mechanisms: (1) activation of the receptor by insulin mimetics that bind in the extracellular domain and (2) inhibition of the receptor TK enzymatic activity in the cytoplasmic domain. While a complete structural picture of the full-length receptor comprising the entire sequence covering extracellular, transmembrane, juxtamembrane and cytoplasmic domains is still elusive, recent progress through cryoelectron microscopy has made it possible to describe the initial insulin ligand binding events at atomistic detail. We utilize this opportunity to obtain structural insights into the pharmacology of the insulin receptor. To this end, we conducted a comprehensive docking study of known ligands to the new structures of the receptor. Through this approach, we provide an in-depth, structure-based review of human insulin receptor pharmacology in light of the new structures. LINKED ARTICLES: This article is part of a themed issue on Structure Guided Pharmacology of Membrane Proteins (BJP 75th Anniversary). To view the other articles in this section visit http://onlinelibrary.wiley.com/doi/10.1111/bph.v179.14/issuetoc., (© 2021 The British Pharmacological Society.)
- Published
- 2022
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36. Identification of allosteric fingerprints of alpha-synuclein aggregates in matrix metalloprotease-1 and substrate-specific virtual screening with single molecule insights.
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Kamboj S, Harms C, Wright D, Nash A, Kumar L, Klein-Seetharaman J, and Sarkar SK
- Subjects
- Catalytic Domain, Molecular Docking Simulation, Protein Aggregation, Pathological metabolism, Matrix Metalloproteinase 1 genetics, Matrix Metalloproteinase 1 metabolism, alpha-Synuclein metabolism
- Abstract
Alpha-synuclein (aSyn) has implications in pathological protein aggregations in neurodegeneration. Matrix metalloproteases (MMPs) are broad-spectrum proteases and cleave aSyn, leading to aggregation. Previous reports showed that allosteric communications between the two domains of MMP1 on collagen fibril and fibrin depend on substrates, activity, and ligands. This paper reports quantification of allostery using single molecule measurements of MMP1 dynamics on aSyn-induced aggregates by calculating Forster Resonance Energy Transfer (FRET) between two dyes attached to the catalytic and hemopexin domains of MMP1. The two domains of MMP1 prefer open conformations that are inhibited by a single point mutation E219Q of MMP1 and tetracycline, an MMP inhibitor. A two-state Poisson process describes the interdomain dynamics, where the two states and kinetic rates of interconversion between them are obtained from histograms and autocorrelations of FRET values. Since a crystal structure of aSyn-bound MMP1 is unavailable, binding poses were predicted by molecular docking of MMP1 with aSyn using ClusPro. MMP1 dynamics were simulated using predicted binding poses and compared with the experimental interdomain dynamics to identify an appropriate pose. The selected aSyn-MMP1 binding pose near aSyn residue K45 was simulated and analyzed to define conformational changes at the catalytic site. Allosteric residues in aSyn-bound MMP1 exhibiting strong correlations with the catalytic motif residues were compared with allosteric residues in free MMP1, and aSyn-specific residues were identified. The allosteric residues in aSyn-bound MMP1 are K281, T283, G292, G327, L328, E329, R337, F343, G345, N346, Y348, G353, Q354, D363, Y365, S366, S367, F368, P371, R372, V374, K375, A379, F391, A394, R399, M414, F419, V426, and C466. Shannon entropy was defined to quantify MMP1 dynamics. Virtual screening was performed against a site on selected aSyn-MMP1 binding poses, which showed that lead molecules differ between free MMP1 and substrate-bound MMP1. Also, identifying aSyn-specific allosteric residues in MMP1 enabled further selection of lead molecules. In other words, virtual screening needs to take substrates into account for potential substrate-specific control of MMP1 activity in the future. Molecular understanding of interactions between MMP1 and aSyn-induced aggregates may open up the possibility of degrading aggregates by targeting MMPs., (© 2022. The Author(s).)
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- 2022
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37. Interpretable network propagation with application to expanding the repertoire of human proteins that interact with SARS-CoV-2.
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Law JN, Akers K, Tasnina N, Santina CMD, Deutsch S, Kshirsagar M, Klein-Seetharaman J, Crovella M, Rajagopalan P, Kasif S, and Murali TM
- Subjects
- Algorithms, Humans, Protein Interaction Maps, Proteins metabolism, COVID-19, SARS-CoV-2
- Abstract
Background: Network propagation has been widely used for nearly 20 years to predict gene functions and phenotypes. Despite the popularity of this approach, little attention has been paid to the question of provenance tracing in this context, e.g., determining how much any experimental observation in the input contributes to the score of every prediction., Results: We design a network propagation framework with 2 novel components and apply it to predict human proteins that directly or indirectly interact with SARS-CoV-2 proteins. First, we trace the provenance of each prediction to its experimentally validated sources, which in our case are human proteins experimentally determined to interact with viral proteins. Second, we design a technique that helps to reduce the manual adjustment of parameters by users. We find that for every top-ranking prediction, the highest contribution to its score arises from a direct neighbor in a human protein-protein interaction network. We further analyze these results to develop functional insights on SARS-CoV-2 that expand on known biology such as the connection between endoplasmic reticulum stress, HSPA5, and anti-clotting agents., Conclusions: We examine how our provenance-tracing method can be generalized to a broad class of network-based algorithms. We provide a useful resource for the SARS-CoV-2 community that implicates many previously undocumented proteins with putative functional relationships to viral infection. This resource includes potential drugs that can be opportunistically repositioned to target these proteins. We also discuss how our overall framework can be extended to other, newly emerging viruses., (© The Author(s) 2021. Published by Oxford University Press GigaScience.)
- Published
- 2021
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38. Similarity of the non-amyloid-β component and C-terminal tail of monomeric and tetrameric alpha-synuclein with 14-3-3 sigma.
- Author
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Evans SR, West C, and Klein-Seetharaman J
- Abstract
Alpha-synuclein (αSyn) is often described as a predominantly disordered protein that has a propensity to self-assemble into toxic oligomers that are found in patients with Parkinson's and Alzheimer's diseases. αSyn's chaperone behavior and tetrameric structure are proposed to be protective against toxic oligomerization. In this paper, we extended the previously proposed similarity between αSyn and 14-3-3 proteins to the α-helical tetrameric species of αSyn in detail. 14-3-3 proteins are a family of well-folded proteins with seven human isoforms, and function in signal transduction and as molecular chaperones. We investigated protein homology, using sequence alignment, amyloid, and disorder prediction, as well as three-dimensional visualization and protein-interaction networks. Our results show sequence homology and structural similarity between the aggregation-prone non-amyloid-β component (NAC) residues Val-52 to Gly-111 in αSyn and 14-3-3 sigma residues Leu-12 to Gly-78. We identified an additional region of sequence homology in the C-terminal region of αSyn (residues Ser-129 to Asp-135) and a C-terminal loop of 14-3-3 between helix αH and αI (residues Ser-209 to Asp-215). This data indicates αSyn shares conserved domain architecture with small heat shock proteins. We show predicted regions of high amyloidogenic propensity and intrinsic structural disorder in αSyn coincide with amyloidogenic and disordered predictions for 14-3-3 proteins. The homology in the NAC region aligns with residues involved in dimer- and tetramerization of the non-amyloidogenic 14-3-3 proteins. Because 14-3-3 proteins are generally not prone to misfolding, our results lend further support to the hypothesis that the NAC region is critical to the assembly of αSyn into the non-toxic tetrameric state., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2021 The Authors.)
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- 2021
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39. Applying model approaches in non-model systems: A review and case study on coral cell culture.
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Roger LM, Reich HG, Lawrence E, Li S, Vizgaudis W, Brenner N, Kumar L, Klein-Seetharaman J, Yang J, Putnam HM, and Lewinski NA
- Subjects
- Animals, Anthozoa growth & development, Cell Culture Techniques methods
- Abstract
Model systems approaches search for commonality in patterns underlying biological diversity and complexity led by common evolutionary paths. The success of the approach does not rest on the species chosen but on the scalability of the model and methods used to develop the model and engage research. Fine-tuning approaches to improve coral cell cultures will provide a robust platform for studying symbiosis breakdown, the calcification mechanism and its disruption, protein interactions, micronutrient transport/exchange, and the toxicity of nanoparticles, among other key biological aspects, with the added advantage of minimizing the ethical conundrum of repeated testing on ecologically threatened organisms. The work presented here aimed to lay the foundation towards development of effective methods to sort and culture reef-building coral cells with the ultimate goal of obtaining immortal cell lines for the study of bleaching, disease and toxicity at the cellular and polyp levels. To achieve this objective, the team conducted a thorough review and tested the available methods (i.e. cell dissociation, isolation, sorting, attachment and proliferation). The most effective and reproducible techniques were combined to consolidate culture methods and generate uncontaminated coral cell cultures for ~7 days (10 days maximum). The tests were conducted on scleractinian corals Pocillopora acuta of the same genotype to harmonize results and reduce variation linked to genetic diversity. The development of cell separation and identification methods in conjunction with further investigations into coral cell-type specific metabolic requirements will allow us to tailor growth media for optimized monocultures as a tool for studying essential reef-building coral traits such as symbiosis, wound healing and calcification at multiple scales., Competing Interests: The authors have declared that no competing interests exist.
- Published
- 2021
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40. Digital image processing to detect subtle motion in stony coral.
- Author
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Li S, Roger LM, Kumar L, Lewinski NA, Klein-Seetharaman J, Gagnon A, Putnam HM, and Yang J
- Abstract
Coral reef ecosystems support significant biological activities and harbor huge diversity, but they are facing a severe crisis driven by anthropogenic activities and climate change. An important behavioral trait of the coral holobiont is coral motion, which may play an essential role in feeding, competition, reproduction, and thus survival and fitness. Therefore, characterizing coral behavior through motion analysis will aid our understanding of basic biological and physical coral functions. However, tissue motion in the stony scleractinian corals that contribute most to coral reef construction are subtle and may be imperceptible to both the human eye and commonly used imaging techniques. Here we propose and apply a systematic approach to quantify and visualize subtle coral motion across a series of light and dark cycles in the scleractinian coral Montipora capricornis. We use digital image correlation and optical flow techniques to quantify and characterize minute coral motions under different light conditions. In addition, as a visualization tool, motion magnification algorithm magnifies coral motions in different frequencies, which explicitly displays the distinctive dynamic modes of coral movement. Specifically, our assessment of displacement, strain, optical flow, and mode shape quantify coral motion under different light conditions, and they all show that M. capricornis exhibits more active motions at night compared to day. Our approach provides an unprecedented insight into micro-scale coral movement and behavior through macro-scale digital imaging, thus offering a useful empirical toolset for the coral research community.
- Published
- 2021
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41. Cephalosporins Interfere With Quorum Sensing and Improve the Ability of Caenorhabditis elegans to Survive Pseudomonas aeruginosa Infection.
- Author
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Kumar L, Brenner N, Brice J, Klein-Seetharaman J, and Sarkar SK
- Abstract
Pseudomonas aeruginosa utilizes the quorum sensing (QS) system to strategically coordinate virulence and biofilm formation. Targeting QS pathways may be a potential anti-infective approach to treat P. aeruginosa infections. In the present study, we define cephalosporins' anti-QS activity using Chromobacterium violaceum CV026 for screening and QS-regulated mutants of P. aeruginosa for validation. We quantified the effects of three cephalosporins, cefepime, ceftazidime, and ceftriaxone, on (1) pyocyanin production using spectrophotometric assay, (2) bacterial motility using agar plate assay, and (3) biofilm formation using scanning electron microscopy. We also studied isogenic QS mutant strains of PAO1 (Δ lasR ,Δ rhlR ,Δ pqsA , and Δ pqsR) to compare and distinguish QS-mediated effects on the motility phenotypes and bacterial growth with and without sub-MIC concentrations of antibiotics. Results showed that cephalosporins have anti-QS activity and reduce bacterial motility, pyocyanin production, and biofilm formation for CV026 and PAO1. Also, sub-MICs of cefepime increased aminoglycosides' antimicrobial activity against P. aeruginosa PAO1, suggesting the advantage of combined anti-QS and antibacterial treatment. To correlate experimentally observed anti-QS effects with the interactions between cephalosporins and QS receptors, we performed molecular docking with ligand binding sites of quorum sensing receptors using Autodock Vina. Molecular docking predicted cephalosporins' binding affinities to the ligand-binding pocket of QS receptors (CviR, LasR, and PqsR). To validate our results using an infection model, we quantified the survival rate of C aenorhabditis elegans following P. aeruginosa PAO1 challenge at concentrations less than the minimum inhibitory concentration (MIC) of antibiotics. C. elegans infected with PAO1 without antibiotics showed 0% survivability after 72 h. In contrast, PAO1-infected C. elegans showed 65 ± 5%, 58 ± 4%, and 49 ± 8% survivability after treatment with cefepime, ceftazidime, and ceftriaxone, respectively. We determined the survival rates of C. elegans infected by QS mutant strains Δ lasR (32 ± 11%), Δ rhlR (27 ± 8%), Δ pqsA (27 ± 10%), and Δ pqsR (37 ± 6%), which suggest essential role of QS system in virulence. In summary, cephalosporins at sub-MIC concentrations show anti-QS activity and enhance the antibacterial efficacy of aminoglycosides, a different class of antibiotics. Thus, cephalosporins at sub-MIC concentrations in combination with other antibiotics are potential candidates for developing therapies to combat infections caused by P. aeruginosa., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Kumar, Brenner, Brice, Klein-Seetharaman and Sarkar.)
- Published
- 2021
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42. A method of purifying alpha-synuclein in E. coli without chromatography.
- Author
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Kamboj S, Harms C, Kumar L, Creamer D, West C, Klein-Seetharaman J, and Sarkar SK
- Abstract
Research has implicated alpha-synuclein (aSyn) in pathological protein aggregation observed in almost all patients with Parkinson's disease and more than 50% of patients with Alzheimer's disease. An easy and inexpensive method of purifying aSyn and developing an in vitro model system of Lewy body formation would enhance basic biomedical research. We report aSyn purification technique that leverages the amyloidogenic property of aSyn suitable for purifying monomeric aSyn without chromatography and denaturing agents. We expressed full-length and untagged aSyn in Rosetta(DE3) pLysS and purified ~60 μg of aSyn from 500 mL culture within 24 h. After IPTG-induced expression of aSyn in E. coli , we disrupted the cells with a sonicator. We centrifuged the cell lysate in a 15 mL tube, which leads to aSyn-induced aggregation of native E. coli proteins. After removing aggregates, centrifugation in a 30 kDa cut-off filter followed by a 10 kDa cut-off filter led to purified water-soluble aSyn. The identity of aSyn was confirmed by Western blot using anti-aSyn antibody and Edman sequencing. Its mass was determined to be 14.6 kDa using a MALDI TOF-MS mass spectrometer. The majority of aSyn led to water-suspended (as opposed to precipitated) aggregation of E. coli proteins with visible fibrous structures. The broad-spectrum binding and amyloidogenic property of aSyn is thus not only useful for inexpensive aSyn production for diverse applications, but it also expands studying its possible roles in human physiology. The aggregate of E. coli proteins induced by aSyn during the purification process may serve as a Lewy body model., Competing Interests: Patent pending., (© 2020 The Author(s).)
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- 2021
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43. Protein sequence models for prediction and comparative analysis of the SARS-CoV-2 -human interactome.
- Author
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Kshirsagar M, Tasnina N, Ward MD, Law JN, Murali TM, Lavista Ferres JM, Bowman GR, and Klein-Seetharaman J
- Subjects
- Amino Acid Sequence, Computational Biology, Humans, Proteins, COVID-19, SARS-CoV-2
- Abstract
Viruses such as the novel coronavirus, SARS-CoV-2, that is wreaking havoc on the world, depend on interactions of its own proteins with those of the human host cells. Relatively small changes in sequence such as between SARS-CoV and SARS-CoV-2 can dramatically change clinical phenotypes of the virus, including transmission rates and severity of the disease. On the other hand, highly dissimilar virus families such as Coronaviridae, Ebola, and HIV have overlap in functions. In this work we aim to analyze the role of protein sequence in the binding of SARS-CoV-2 virus proteins towards human proteins and compare it to that of the above other viruses. We build supervised machine learning models, using Generalized Additive Models to predict interactions based on sequence features and find that our models perform well with an AUC-PR of 0.65 in a class-skew of 1:10. Analysis of the novel predictions using an independent dataset showed statistically significant enrichment. We further map the importance of specific amino-acid sequence features in predicting binding and summarize what combinations of sequences from the virus and the host is correlated with an interaction. By analyzing the sequence-based embeddings of the interactomes from different viruses and clustering them together we find some functionally similar proteins from different viruses. For example, vif protein from HIV-1, vp24 from Ebola and orf3b from SARS-CoV all function as interferon antagonists. Furthermore, we can differentiate the functions of similar viruses, for example orf3a's interactions are more diverged than orf7b interactions when comparing SARS-CoV and SARS-CoV-2.
- Published
- 2021
44. Bioinformatics of corals: Investigating heterogeneous omics data from coral holobionts for insight into reef health and resillience.
- Author
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Cowen LJ, Klein-Seetharaman J, and Putnam H
- Subjects
- Animals, Computational Biology, Coral Reefs, Anthozoa genetics, Microbiota
- Abstract
Coral reefs are home to over 2 million species and provide habitat for roughly 25% of all marine animals, but they are being severely threatened by pollution and climate change. A large amount of genomic, transcriptomic and other -omics data from different species of reef building corals, the uni-cellular dinoagellates, plus the coral microbiome (where corals have possibly the most complex microbiome yet discovered, consisting of over 20,000 different species), is becoming increasingly available for corals. This new data present an opportunity for bioinformatics researchers and computational biologists to contribute to a timely, compelling, and urgent investigation of critical factors that influence reef health and resilience. This paper summarizes the content of the Bioinformatics of Corals workshop, that is being held as part of PSB 2021. It is particularly relevant for this workshop to occur at PSB, given the abundance of and reliance on coral reefs in Hawaii and the conference's traditional association with the region.
- Published
- 2021
45. Activity-dependent interdomain dynamics of matrix metalloprotease-1 on fibrin.
- Author
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Kumar L, Planas-Iglesias J, Harms C, Kamboj S, Wright D, Klein-Seetharaman J, and Sarkar SK
- Subjects
- Catalysis, Catalytic Domain physiology, Escherichia coli metabolism, Fibrinogen metabolism, Hemopexin metabolism, Kinetics, Matrix Metalloproteinase 1 metabolism, Protein Domains physiology
- Abstract
The roles of protein conformational dynamics and allostery in function are well-known. However, the roles that interdomain dynamics have in function are not entirely understood. We used matrix metalloprotease-1 (MMP1) as a model system to study the relationship between interdomain dynamics and activity because MMP1 has diverse substrates. Here we focus on fibrin, the primary component of a blood clot. Water-soluble fibrinogen, following cleavage by thrombin, self-polymerize to form water-insoluble fibrin. We studied the interdomain dynamics of MMP1 on fibrin without crosslinks using single-molecule Forster Resonance Energy Transfer (smFRET). We observed that the distance between the catalytic and hemopexin domains of MMP1 increases or decreases as the MMP1 activity increases or decreases, respectively. We modulated the activity using (1) an active site mutant (E219Q) of MMP1, (2) MMP9, another member of the MMP family that increases the activity of MMP1, and (3) tetracycline, an inhibitor of MMP1. We fitted the histograms of smFRET values to a sum of two Gaussians and the autocorrelations to an exponential and power law. We modeled the dynamics as a two-state Poisson process and calculated the kinetic rates from the histograms and autocorrelations. Activity-dependent interdomain dynamics may enable allosteric control of the MMP1 function.
- Published
- 2020
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46. Insights into the Mechanism of Action of Highly Diluted Biologics.
- Author
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Tarasov SA, Gorbunov EA, Don ES, Emelyanova AG, Kovalchuk AL, Yanamala N, Schleker ASS, Klein-Seetharaman J, Groenestein R, Tafani JP, van der Meide P, and Epstein OI
- Subjects
- Amino Acids metabolism, Animals, Cell Line, Cell Line, Tumor, Dogs, Female, Humans, Influenza A virus drug effects, Interferon-gamma metabolism, Leukocytes, Mononuclear drug effects, Madin Darby Canine Kidney Cells, Mice, Mice, Inbred BALB C, Orthomyxoviridae Infections drug therapy, Orthomyxoviridae Infections metabolism, U937 Cells, Biological Products pharmacology
- Abstract
The therapeutic use of Abs in cancer, autoimmunity, transplantation, and other fields is among the major biopharmaceutical advances of the 20th century. Broader use of Ab-based drugs is constrained because of their high production costs and frequent side effects. One promising approach to overcome these limitations is the use of highly diluted Abs, which are produced by gradual reduction of an Ab concentration to an extremely low level. This technology was used to create a group of drugs for the treatment of various diseases, depending on the specificity of the used Abs. Highly diluted Abs to IFN-γ (hd-anti-IFN-γ) have been demonstrated to be efficacious against influenza and other respiratory infections in a variety of preclinical and clinical studies. In the current study, we provide evidence for a possible mechanism of action of hd-anti-IFN-γ. Using high-resolution solution nuclear magnetic resonance spectroscopy, we show that the drug induced conformational changes in the IFN-γ molecule. Chemical shift changes occurred in the amino acids located primarily at the dimer interface and at the C-terminal region of IFN-γ. These molecular changes could be crucial for the function of the protein, as evidenced by an observed hd-anti-IFN-γ-induced increase in the specific binding of IFN-γ to its receptor in U937 cells, enhanced induced production of IFN-γ in human PBMC culture, and increased survival of influenza A-infected mice., (Copyright © 2020 by The American Association of Immunologists, Inc.)
- Published
- 2020
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47. Lemon Extract Reduces Angiotensin Converting Enzyme (ACE) Expression and Activity and Increases Insulin Sensitivity and Lipolysis in Mouse Adipocytes.
- Author
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Tejpal S, Wemyss AM, Bastie CC, and Klein-Seetharaman J
- Subjects
- 3T3-L1 Cells, Adipocytes drug effects, Adipose Tissue, Animals, Cell Differentiation drug effects, Disease Models, Animal, Glucose Transporter Type 4 metabolism, Insulin blood, Lipid Metabolism drug effects, Mice, Phosphorylation, Proto-Oncogene Proteins c-akt metabolism, Sterol Esterase metabolism, Angiotensin-Converting Enzyme Inhibitors pharmacology, Citrus, Insulin Resistance physiology, Lipolysis drug effects, Peptidyl-Dipeptidase A drug effects, Plant Extracts pharmacology
- Abstract
Obesity is associated with insulin resistance and cardiovascular complications. In this paper, we examine the possible beneficial role of lemon juice in dieting. Lemon extract (LE) has been proposed to improve serum insulin levels and decrease angiotensin converting enzyme (ACE) activity in mouse models. ACE is also a biomarker for sustained weight loss and ACE inhibitors improve insulin sensitivity in humans. Here, we show that LE impacts adipose tissue metabolism directly. In 3T3-L1 differentiated adipocyte cells, LE improved insulin sensitivity as evidenced by a 3.74 ± 0.54-fold increase in both pAKT and GLUT4 levels. LE also induced lipolysis as demonstrated by a 16.6 ± 1.2 fold-change in pHSL protein expression levels. ACE gene expression increased 12.0 ± 0.1 fold during differentiation of 3T3-L1 cells in the absence of LE, and treatment with LE decreased ACE gene expression by 80.1 ± 0.5% and protein expression by 55 ± 0.37%. We conclude that LE's reduction of ACE expression causes increased insulin sensitivity and breakdown of lipids in adipocytes.
- Published
- 2020
- Full Text
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48. Allosteric Communications between Domains Modulate the Activity of Matrix Metalloprotease-1.
- Author
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Kumar L, Nash A, Harms C, Planas-Iglesias J, Wright D, Klein-Seetharaman J, and Sarkar SK
- Subjects
- Catalytic Domain, Kinetics, Proteins, Matrix Metalloproteinase 1 genetics, Matrix Metalloproteinase 1 metabolism, Molecular Dynamics Simulation
- Abstract
An understanding of the structure-dynamics relationship is essential for understanding how a protein works. Prior research has shown that the activity of a protein correlates with intradomain dynamics occurring at picosecond to millisecond timescales. However, the correlation between interdomain dynamics and the function of a protein is poorly understood. Here, we show that communications between the catalytic and hemopexin domains of matrix metalloprotease-1 (MMP1) on type 1 collagen fibrils correlate with its activity. Using single-molecule Förster resonance energy transfer, we identified functionally relevant open conformations in which the two MMP1 domains are well separated, which were significantly absent for catalytically inactive point mutant (E219Q) of MMP1 and could be modulated by an inhibitor or an enhancer of activity. The observed relevance of open conformations resolves the debate about the roles of open and closed MMP1 structures in function. We fitted the histograms of single-molecule Förster resonance energy transfer values to a sum of two Gaussians and the autocorrelations to an exponential and power law. We used a two-state Poisson process to describe the dynamics and calculate the kinetic rates from the fit parameters. All-atom and coarse-grained simulations reproduced some of the experimental features and revealed substrate-dependent MMP1 dynamics. Our results suggest that an interdomain separation facilitates opening up the catalytic pocket so that the collagen chains come closer to the MMP1 active site. Coordination of functional conformations at different parts of MMP1 occurs via allosteric communications that can take place via interactions mediated by collagen even if the linker between the domains is absent. Modeling dynamics as a Poisson process enables connecting the picosecond timescales of molecular dynamics simulations with the millisecond timescales of single-molecule measurements. Water-soluble MMP1 interacting with water-insoluble collagen fibrils poses challenges for biochemical studies that the single-molecule tracking can overcome for other insoluble substrates. Interdomain communications are likely important for multidomain proteins., (Copyright © 2020 Biophysical Society. Published by Elsevier Inc. All rights reserved.)
- Published
- 2020
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49. Angiotensin Converting Enzyme (ACE): A Marker for Personalized Feedback on Dieting.
- Author
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Tejpal S, Sanghera N, Manoharan V, Planas-Iglesias J, Bastie CC, and Klein-Seetharaman J
- Subjects
- Biomarkers metabolism, Body Mass Index, Humans, Insulin metabolism, Lactic Acid metabolism, Life Style, Weight Loss, Diet, Feedback, Peptidyl-Dipeptidase A metabolism
- Abstract
Angiotensin Converting Enzyme (ACE) expression and activity is associated with obesity. ACE is a circulating factor that predicts sustained weight loss over a time frame of months. Here, we evaluate whether ACE might also be an early marker (over a 24-hour period) for weight loss. 32 participants (78% females; BMI 28.47 ± 4.87kg/m2) followed a 1200KCal diet with an optional daily (<250KCal) snack and were asked to use an in-house generated health platform to provide recordings of food intake, physical activity and urine collection time and volume. Following a day of dieting, ACE levels in urine negatively correlated with weight loss ( p = 0.015 ). This reduction in ACE levels was significantly more robust in individuals with a BMI > 25 ( p = 0.0025 ). This study demonstrated that ACE levels correlate with BMI and weight loss as early as after 1 day of dieting, and thus ACE could be a potential early "biofeedback" marker for weight loss and diet efficiency., Competing Interests: “The authors declare no conflict of interest.”
- Published
- 2020
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50. Correction: Comparison of the molecular properties of retinitis pigmentosa P23H and N15S amino acid replacements in rhodopsin.
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Mitchell J, Balem F, Tirupula K, Man D, Dhiman HK, Yanamala N, Ollesch J, Planas-Iglesias J, Jennings BJ, Gerwert K, Iannaccone A, and Klein-Seetharaman J
- Abstract
[This corrects the article DOI: 10.1371/journal.pone.0214639.].
- Published
- 2019
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- View/download PDF
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