720 results on '"Daly, Norelle L."'
Search Results
302. IgE and IgG4 epitopes revealed on the major fish allergen Lat c 1.
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Sharp, Michael F., Taki, Aya C., Ruethers, Thimo, Stephen, Juan N., Daly, Norelle L., Lopata, Andreas L., and Kamath, Sandip D.
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LATEX , *ALLERGENS , *EPITOPES , *AMINO acid sequence , *FISHES , *BINDING sites , *CALBINDIN , *IMMUNOGLOBULIN G - Abstract
• The IgE- and IgG 4 -binding to major fish allergen Lat c 1.01 was investigated. • The IgG 4 binding was patient-specific regardless of the age group. • Two specific IgE epitopes were identified at the C- and N-terminal regions. • Highlighted the importance of epitope analysis for fish allergy management. The IgE- and IgG 4 -binding patterns of the major fish allergen parvalbumins are not clearly understood. IgE antibody-binding to parvalbumin from Asian seabass, Lat c 1.01, is implicated in up to 90 % of allergic reactions, although the region of IgE or IgG4 epitopes are unknown. In the present study, we characterized the specific IgE- and IgG 4 -binding regions of Lat c 1.01 using serum from pediatric and adult patients with clinically-confirmed fish allergy. A comparative investigation of patient IgE- and IgG 4 -binding to recombinant Lat c 1.01 was performed by immunoblotting and indirect ELISA using serum from 15 children and eight adults with clinically confirmed IgE-mediated reactions to fish. The IgE- and IgG 4 -binding regions of Lat c 1.01 were determined by inhibition ELISA using seven overlapping peptides spanning the entire 102 amino acid sequence. Elucidated IgE-binding regions were modelled and compared to known antibody-binding regions of parvalbumins from five other fish species. Ninety five percent (22/23) patients demonstrated IgE-binding to rLat c 1.01, while fewer patients (10/15 children and 7/8 adults) demonstrated robust IgG 4 binding when determined by immunoblots. IgE-binding for both cohorts was significantly higher compared to IgG 4 -binding by ELISA. All patients in this study presented individual IgE and IgG 4 epitope-recognition profiles. In addition to these patient-specific antibody binding sites, general IgE epitopes were also identified at the C- and N-terminal regions of this major fish allergen. Our findings demonstrate two specific IgE epitopes on parvalbumin from Asian seabass, while IgG 4 binding is much lower and patient specific. This study highlights the importance of advancement in epitope analysis regardless of the age group for diagnostics and immunotherapies for fish allergy. [ABSTRACT FROM AUTHOR]
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- 2021
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303. Structure–activity relationship and conformational studies of the natural product cyclic depsipeptides YM-254890 and FR900359.
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Zhang, Hang, Nielsen, Alexander L., Boesgaard, Michael W., Harpsøe, Kasper, Daly, Norelle L., Xiong, Xiao-Feng, Underwood, Christina R., Haugaard-Kedström, Linda M., Bräuner-Osborne, Hans, Gloriam, David E., and Strømgaard, Kristian
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PEPTIDES , *SPECTRUM analysis , *STEREOCHEMISTRY , *PROTEINS , *OPTICS - Abstract
G proteins are key mediators in the signaling of G protein-coupled receptors and involved in a plethora of important physiological processes. The natural product cyclic depsipeptides YM-254890 and FR900359 are the only known selective inhibitors of the G q protein subfamily. So far, all reported YM-254890 and FR900359 analogs show no inhibition of other G protein subtypes except the G q , G 11 and G 14 proteins. Here we report the rationalization of the high potency of FR900359 and efforts towards understanding the G protein subtype selectivity by synthesis of a collection of structurally and stereochemically diverse analogs of YM-254890 using an efficient synthetic protocol. We performed the first conformational study of YM-254890 in aqueous solution by NMR spectroscopy and replica exchange molecular dynamics, which suggested that the combined contribution of residues with appropriate size, stereochemistry and conformational stability are critical for inhibitory potency. Moreover, in addition to the fit of the binding pocket, more factors should be taken into consideration for the development of compounds targeting other G proteins. [ABSTRACT FROM AUTHOR]
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- 2018
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304. Development of Novel Melanocortin Receptor Agonists Based on the Cyclic Peptide Framework of Sunflower Trypsin Inhibitor-1.
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Durek, Thomas, Cromm, Philipp M., White, Andrew M., Schroeder, Christina I., Kaas, Quentin, Weidmann, Joachim, Fuaad, Abdullah Ahmad, Cheneval, Olivier, Harvey, Peta J., Daly, Norelle L., Yang Zhou, Dellsén, Anita, Österlund, Torben, Larsson, Niklas, Knerr, Laurent, Bauer, Udo, Kessler, Horst, Cai, Minying, Hruby, Victor J., and Plowright, Alleyn T.
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MELANOCORTIN receptors , *TRYPSIN inhibitors - Abstract
Ultrastable cyclic peptide frameworks offer great potential for drug design due to their improved bioavailability compared to their linear analogues. Using the sunflower trypsin inhibitor-1 (SFTI-1) peptide scaffold in combination with systematic N-methylation of the grafted pharmacophore led to the identification of novel subtype selective melanocortin receptor (MCR) agonists. Multiple bicyclic peptides were synthesized and tested toward their activity at MC1R and MC3–5R. Double N-methylated compound 18 showed a pKi of 8.73 ± 0.08 (Ki = 1.92 ± 0.34 nM) and a pEC50 of 9.13 ± 0.04 (EC50 = 0.75 ± 0.08 nM) at the human MC1R and was over 100 times more selective for MC1R. Nuclear magnetic resonance structural analysis of 18 emphasized the role of peptide bond N-methylation in shaping the conformation of the grafted pharmacophore. More broadly, this study highlights the potential of cyclic peptide scaffolds for epitope grafting in combination with N-methylation to introduce receptor subtype selectivity in the context of peptide-based drug discovery. [ABSTRACT FROM AUTHOR]
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- 2018
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305. Conotoxin Φ-MiXXVIIA from the Superfamily G2 Employs a Novel Cysteine Framework that Mimics Granulin and Displays Anti-Apoptotic Activity.
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Jin, Ai ‐ Hua, Dekan, Zoltan, Smout, Michael J., Wilson, David, Dutertre, Sébastien, Vetter, Irina, Lewis, Richard J., Loukas, Alex, Daly, Norelle L., and Alewood, Paul F.
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CYSTEINE , *DISULFIDES , *CONOTOXINS , *NEUROTOXIC agents , *NUCLEAR magnetic resonance spectroscopy - Abstract
Conotoxins are a large family of disulfide-rich peptides that contain unique cysteine frameworks that target a broad range of ion channels and receptors. We recently discovered the 33-residue conotoxin Φ-MiXXVIIA from Conus miles with a novel cysteine framework comprising three consecutive cysteine residues and four disulfide bonds. Regioselective chemical synthesis helped decipher the disulfide bond connectivity and the structure of Φ-MiXXVIIA was determined by NMR spectroscopy. The 3D structure displays a unique topology containing two β-hairpins that resemble the N-terminal domain of granulin. Similar to granulin, Φ-MiXXVIIA promotes cell proliferation (EC50 17.85 μ m) while inhibiting apoptosis (EC50 2.2 μ m). Additional framework XXVII sequences were discovered with homologous signal peptides that define the new conotoxin superfamily G2. The novel structure and biological activity of Φ-MiXXVIIA expands the repertoire of disulfide-rich conotoxins that recognize mammalian receptors. [ABSTRACT FROM AUTHOR]
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- 2017
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306. The Aromatic Head Group of Spider Toxin Polyamines Influences Toxicity to Cancer Cells.
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Wilson, David, Boyle, Glen M., McIntyre, Lachlan, Nolan, Matthew J., Parsons, Peter G., Smith, Jennifer J., Tribolet, Leon, Loukas, Alex, Liddell, Michael J., Rash, Lachlan D., and Daly, Norelle L.
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SPIDER venom , *CELL-mediated cytotoxicity , *POLYAMINES , *CANCER cells , *NUCLEAR magnetic resonance - Abstract
Spider venoms constitute incredibly diverse libraries of compounds, many of which are involved in prey capture and defence. Polyamines are often prevalent in the venom and target ionotropic glutamate receptors. Here we show that a novel spider polyamine, PA366, containing a hydroxyphenyl-based structure is present in the venom of several species of tarantula, and has selective toxicity against MCF-7 breast cancer cells. By contrast, a polyamine from an Australian funnel-web spider venom, which contains an identical polyamine tail to PA366 but an indole-based head-group, is only cytotoxic at high concentrations. Our results suggest that the ring structure plays a role in the cytotoxicity and that modification to the polyamine head group might lead to more potent and selective compounds with potential as novel cancer treatments. [ABSTRACT FROM AUTHOR]
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- 2017
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307. An engineered cyclic peptide alleviates symptoms of inflammation in a murine model of inflammatory bowel disease.
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Caceres, Claudia Cobos, Bansal, Paramjit S., Navarro, Severine, Wilson, David, Don, Laurianne, Giacomin, Paul, Loukas, Alex, and Daly, Norelle L.
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INFLAMMATORY bowel disease treatment , *PROTEIN engineering , *CYCLIC peptides , *ANNEXINS , *BIOACTIVE compounds - Abstract
Inflammatory bowel diseases (IBDs) are a set of complex and debilitating diseases for which there is no satisfactory treatment. Recent studies have shown that small peptides show promise for reducing inflammation in models of IBD. However, these small peptides are likely to be unstable and rapidly cleared from the circulation, and therefore, if not modified for better stability, represent non-viable drug leads. We hypothesized that improving the stability of these peptides by grafting them into a stable cyclic peptide scaffold may enhance their therapeutic potential. Using this approach, we have designed a novel cyclic peptide that comprises a small bioactive peptide from the annexin A1 protein grafted into a sunflower trypsin inhibitor cyclic scaffold. We used native chemical ligation to synthesize the grafted cyclic peptide. This engineered cyclic peptide maintained the overall fold of the naturally occurring cyclic peptide, was more effective at reducing inflammation in a mouse model of acute colitis than the bioactive peptide alone, and showed enhanced stability in human serum. Our findings suggest that the use of cyclic peptides as structural backbones offers a promising approach for the treatment of IBD and potentially other chronic inflammatory conditions. [ABSTRACT FROM AUTHOR]
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- 2017
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308. Development of a Potent Wound Healing Agent Based on the Liver Fluke Granulin Structural Fold.
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Bansal, Paramjit S., Smout, Michael J., Wilson, David, Cobos Caceres, Claudia, Dastpeyman, Mohadeseh, Sotillo, Javier, Seifert, Julia, Brindley, Paul J., Loukas, Alex, and Daly, Norelle L.
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GROWTH factors , *CELL proliferation , *PARASITIC diseases , *LIVER flukes , *OPISTHORCHIS viverrini - Abstract
Granulins are a family of protein growth factors that are involved in cell proliferation. An orthologue of granulin from the human parasitic liver fluke Opisthorchis viverrini, known as Ov-GRN-1, induces angiogenesis and accelerates wound repair. Recombinant Ov-GRN-1 production is complex and poses an obstacle for clinical development. To identify the bioactive region(s) of Ov-GRN-1, four truncated N-terminal analogues were synthesized and characterized structurally using NMR spectroscopy. Peptides that contained only two native disulfide bonds lack the characteristic granulin β-hairpin structure. Remarkably, the introduction of a non-native disulfide bond was critical for formation of β-hairpin structure. Despite this structural difference, both two and three disulfide-bonded peptides drove proliferation of a human cholangiocyte cell line and demonstrated potent wound healing in mice. Peptides derived from Ov-GRN-1 are leads for novel wound healing therapeutics, as they are likely less immunogenic than the full-length protein and more convenient to produce. [ABSTRACT FROM AUTHOR]
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- 2017
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309. Anti-inflammatory properties of novel galloyl glucosides isolated from the Australian tropical plant Uromyrtus metrosideros.
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Ritmejerytė, Edita, Ryan, Rachael Y.M., Byatt, Brendan J., Peck, Yoshimi, Yeshi, Karma, Daly, Norelle L., Zhao, Guangzu, Crayn, Darren, Loukas, Alex, Pyne, Stephen G., Ruscher, Roland, and Wangchuk, Phurpa
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MONONUCLEAR leukocytes , *GLUCOSIDES , *INFLAMMATORY bowel diseases , *TROPICAL plants , *GLYCOSIDES , *TUMOR necrosis factors , *NUCLEAR magnetic resonance - Abstract
Two new galloyl glucosides, galloyl-lawsoniaside A (4) and uromyrtoside (6), were isolated from the polar fraction of Uromyrtus metrosideros leaf extract along with another four previously identified phytochemicals (1 , 2 , 3 , and 5). The structures of these six compounds were characterised using low and high-resolution mass spectrometry (L/HRMS) and 1D and 2D Nuclear Magnetic Resonance (NMR) spectroscopy. These compounds were not toxic to human peripheral blood mononuclear cells (PBMCs) at 10 μg/mL over 24 h, yet showed significant in vitro suppression of proinflammatory cytokines involved in the pathogenesis of inflammatory bowel disease (IBD). Specifically, the release of interferon γ (IFN-γ), interleukin (IL)-17A, and IL-8 from phorbol myristate acetate/ionomycin (P/I) and anti-CD3/anti-CD28-activated cells were significantly suppressed by compounds 4 and 5. Interestingly, no effect on tumour necrosis factor (TNF) release was observed. These results show that the newly characterised compound 4 has promising cytokine suppressive properties, which could be further investigated as a candidate for IBD treatment. [Display omitted] • Isolated six compounds in total from Uromyrtus metrosideros. • Discovered two new galloyl glucosides-galloyl-lawsoniaside A and uromyrtoside. • These compounds were not toxic to human peripheral blood mononuclear cells. • Galloyl-lawsoniaside A and (4S)-α-terpineol 8- O -β-D-(6- O -galloyl)glucopyranoside showed significant anti-inflammatory activities. [ABSTRACT FROM AUTHOR]
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- 2022
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310. α-conotoxin MrIC is a biased agonist at α7 nicotinic acetylcholine receptors.
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Mueller, Alexander, Starobova, Hana, Inserra, Marco C., Jin, Ai-Hua, Deuis, Jennifer R., Dutertre, Sébastien, Lewis, Richard J., Alewood, Paul F., Daly, Norelle L., and Vetter, Irina
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CONOTOXINS , *NICOTINIC agonists , *NICOTINIC acetylcholine receptors , *PHARMACOLOGY , *ETHYLENEDIAMINETETRAACETIC acid , *ALLOSTERIC regulation - Abstract
MrIC is a recently described selective agonist of endogenously expressed α7 nAChR. In this study, we further characterize the pharmacological activity of MrIC using Ca 2+ imaging approaches in SH-SY5Y cells endogenously expressing α7 nAChR and demonstrate that MrIC exclusively activates α7 nAChR modulated by type II positive allosteric modulators, including PNU120596. MrIC was a full agonist at PNU120596-modulated α7 nAChR compared with choline, albeit with slower kinetics, but failed to elicit a Ca 2+ response in the absence of PNU120596. Interestingly, the NMR structure of MrIC showed a typical 4/7 α-conotoxin fold, indicating that its unusual pharmacological activity is likely sequence-dependent. Overall, our results suggest that MrIC acts as a biased agonist that can only activate α7 nAChR modified by type II positive allosteric modulators, and thus represents a valuable tool to probe the pharmacological properties of this important ion channel. [ABSTRACT FROM AUTHOR]
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- 2015
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311. Solution Structure, Membrane Interactions, and Protein Binding Partners of the Tetraspanin Sm-TSP-2, a Vaccine Antigen from the Human Blood Fluke Schistosoma mansoni.
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Xinying Jia, Schulte, Leigh, Loukas, Alex, Pickering, Darren, Pearson, Mark, Mobli, Mehdi, Jones, Alun, Rosengren, Karl J., Daly, Norelle L., Gobert, Geoffrey N., Jones, Malcolm K., Craik, David J., and Mulvenna, Jason
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TETRASPANIN , *EUKARYOTIC cells , *CELL membranes , *SCHISTOSOMIASIS , *LIPIDS - Abstract
The tetraspanins (TSPs) are a family of integral membrane proteins that are ubiquitously expressed at the surface of eukaryotic cells. TSPs mediate a range of processes at the surface of the plasma membrane by providing a scaffold for the assembly of protein complexes known as tetraspanin-enriched microdomains (TEMs). We report here the structure of the surface- exposed EC2 domain from Sm-TSP-2, a TSP from Schistosoma mansoni and one of the better prospects for the development of a vaccine against schistosomiasis. This is the first solution structure of this domain, and our investigations of its interactions with lipid micelles provide a general model for interactions between TSPs, membranes, and other proteins. Using chemical cross-linking, eight potential protein constituents of Sm-TSP-2-mediated TEMs were also identified. These include proteins important for membrane maintenance and repair, providing further evidence for the functional role of Sm-TSP-2- and Sm-TSP-2-mediated TEMs. The identification of calpain, Sm29, and fructose-bisphosphate aldolase, themselves potential vaccine antigens, suggests that the Sm-TSP-2- mediated TEMs could be disrupted via multiple targets. The identification of further Sm-TSP-2-mediated TEM proteins increases the available candidates for multiplex vaccines and/or novel drugs targeting TEMs in the schistosome tegument. [ABSTRACT FROM AUTHOR]
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- 2014
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312. A Tarantula-Venom Peptide Antagonizes the TRPA1 Nociceptor Ion Channel by Binding to the S1–S4 Gating Domain.
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Gui, Junhong, Liu, Boyi, Cao, Guan, Lipchik, Andrew?M., Perez, Minervo, Dekan, Zoltan, Mobli, Mehdi, Daly, Norelle?L., Alewood, Paul?F., Parker, Laurie?L., King, Glenn?F., Zhou, Yufeng, Jordt, Sven-Eric, and Nitabach, Michael?N.
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TARANTULAS , *PEPTIDES , *ANTIBIOSIS , *ANTISENSE DNA , *ION channels , *NOCICEPTORS ,PHYSIOLOGICAL effects of venom - Abstract
Summary: Background: The venoms of predators have been an excellent source of diverse highly specific peptides targeting ion channels. Here we describe the first known peptide antagonist of the nociceptor ion channel transient receptor potential ankyrin 1 (TRPA1). Results: We constructed a recombinant cDNA library encoding ∼100 diverse GPI-anchored peptide toxins (t-toxins) derived from spider venoms and screened this library by coexpression in Xenopus oocytes with TRPA1. This screen resulted in identification of protoxin-I (ProTx-I), a 35-residue peptide from the venom of the Peruvian green-velvet tarantula, Thrixopelma pruriens, as the first known high-affinity peptide TRPA1 antagonist. ProTx-I was previously identified as an antagonist of voltage-gated sodium (NaV) channels. We constructed a t-toxin library of ProTx-I alanine-scanning mutants and screened this library against NaV1.2 and TRPA1. This revealed distinct partially overlapping surfaces of ProTx-I by which it binds to these two ion channels. Importantly, this mutagenesis yielded two novel ProTx-I variants that are only active against either TRPA1or NaV1.2. By testing its activity against chimeric channels, we identified the extracellular loops of the TRPA1 S1–S4 gating domain as the ProTx-I binding site. Conclusions: These studies establish our approach, which we term “toxineering,” as a generally applicable method for isolation of novel ion channel modifiers and design of ion channel modifiers with altered specificity. They also suggest that ProTx-I will be a valuable pharmacological reagent for addressing biophysical mechanisms of TRPA1 gating and the physiology of TRPA1 function in nociceptors, as well as for potential clinical application in the context of pain and inflammation. [Copyright &y& Elsevier]
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- 2014
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313. Design and Synthesis of Truncated EGF-A Peptides that Restore LDL-R Recycling in the Presence of PCSK9 In Vitro.
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Schroeder, Christina?I., Swedberg, Joakim?E., Withka, Jane?M., Rosengren, K.?Johan, Akcan, Muharrem, Clayton, Daniel?J., Daly, Norelle?L., Cheneval, Olivier, Borzilleri, Kris?A., Griffor, Matt, Stock, Ingrid, Colless, Barbara, Walsh, Phillip, Sunderland, Philip, Reyes, Allan, Dullea, Robert, Ammirati, Mark, Liu, Shenping, McClure, Kim?F., and Tu, Meihua
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EPIDERMAL growth factor , *LOW density lipoproteins , *LIPOPROTEIN receptors , *PROPROTEIN convertases , *SUBTILISINS , *IN vitro studies - Abstract
Summary: Disrupting the binding interaction between proprotein convertase (PCSK9) and the epidermal growth factor-like domain A (EGF-A domain) in the low-density lipoprotein receptor (LDL-R) is a promising strategy to promote LDL-R recycling and thereby lower circulating cholesterol levels. In this study, truncated 26 amino acid EGF-A analogs were designed and synthesized, and their structures were analyzed in solution and in complex with PCSK9. The most potent peptide had an increased binding affinity for PCSK9 (KD = 0.6 μM) compared with wild-type EGF-A (KD = 1.2 μM), and the ability to increase LDL-R recycling in the presence of PCSK9 in a cell-based assay. [Copyright &y& Elsevier]
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- 2014
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314. The C-terminal propeptide of a plant defensin confers cytoprotective and subcellular targeting functions.
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Lay, Fung T., Poon, Simon, McKenna, James A., Connelly, Angela A., Barbeta, Barbara L., McGinness, Bruce S., Fox, Jennifer L., Daly, Norelle L., Craik, David J., Heath, Robyn L., and Anderson, Marilyn A.
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DEFENSINS , *TRANSGENIC plants , *PLANT diseases , *PLANT fibers , *FLORAL products - Abstract
Background Plant defensins are small (45-54 amino acids), basic, cysteine-rich proteins that have a major role in innate immunity in plants. Many defensins are potent antifungal molecules and are being evaluated for their potential to create crop plants with sustainable disease resistance. Defensins are produced as precursor molecules which are directed into the secretory pathway and are divided into two classes based on the absence (class I) or presence (class II) of an acidic C-terminal propeptide (CTPP) of about 33 amino acids. The function of this CTPP had not been defined. Results By transgenically expressing the class II plant defensin NaD1 with and without its cognate CTPP we have demonstrated that NaD1 is phytotoxic to cotton plants when expressed without its CTPP. Transgenic cotton plants expressing constructs encoding the NaD1 precursor with the CTPP had the same morphology as non-transgenic plants but expression of NaD1 without the CTPP led to plants that were stunted, had crinkled leaves and were less viable. Immunofluorescence microscopy and transient expression of a green fluorescent protein (GFP)-CTPP chimera were used to confirm that the CTPP is sufficient for vacuolar targeting. Finally circular dichroism and NMR spectroscopy were used to show that the CTPP adopts a helical confirmation. Conclusions In this report we have described the role of the CTPP on NaD1, a class II defensin from Nicotiana alata flowers. The CTPP of NaD1 is sufficient for vacuolar targeting and plays an important role in detoxification of the defensin as it moves through the plant secretory pathway. This work may have important implications for the use of defensins for disease protection in transgenic crops. [ABSTRACT FROM AUTHOR]
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- 2014
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315. Oxytocic plant cyclotides as templates for peptide G protein-coupled receptor ligand design.
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Koehbach, Johannes, O'Brien, Margaret, Muttenthaler, Markus, Miazzo, Marion, Akcan, Muharrem, Elliott, Alysha G., Daly, Norelle L., Harvey, Peta J., Arrowsmith, Sarah, Gunasekera, Sunithi, Smith, Terry J., Wray, Susan, Göransson, Ulf, Dawson, Philip E., Craik, David J., Freissmuth, Michael, and Gruber, Christian W.
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G proteins , *PITUITARY hormones , *OXYTOCIN , *RUBIACEAE , *MUSCLE cells - Abstract
Cyclotides are plant peptides comprising a circular backbone and three conserved disulfide bonds that confer them with exceptional stability. They were originally discovered in Oldenlandia affinis based on their use in traditional African medicine to accelerate labor. Recently, cyclotides have been identified in numerous plant species of the coffee, violet, cucurbit, pea, potato, and grass families. Their unique structural topology, high stability, and tolerance to sequence variation make them promising templates for the development of peptide-based pharmaceuticals. However, the mechanisms underlying their biological activities remain largely unknown; specifically, a receptor for a native cyclotide has not been reported hitherto. Using bioactivity-guided fractionation of an herbal peptide extract known to indigenous healers as "kalata-kalata," the cyclotide kalata B7 was found to induce strong contractility on human uterine smooth muscle cells. Radioligand displacement and second messenger-based reporter assays confirmed the oxytocin and vasopressin V1a receptors, members of the G protein-coupled receptor family, as molecular targets for this cyclotide. Furthermore, we show that cyclotides can serve as templates for the design of selective G protein-coupled receptor ligands by generating an oxytocin-like peptide with nanomolar affinity. This nonapeptide elicited dose-dependent contractions on human myometrium. These observations provide a proof of concept for the development of cyclotide-based peptide ligands. [ABSTRACT FROM AUTHOR]
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- 2013
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316. Novel Inhibitor Cystine Knot Peptides from Momordica charantia.
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He, Wen-Jun, Chan, Lai Yue, Clark, Richard J., Tang, Jun, Zeng, Guang-Zhi, Franco, Octavio L., Cantacessi, Cinzia, Craik, David J., Daly, Norelle L., and Tan, Ning-Hua
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CYSTINE , *ENZYME inhibitors , *MOMORDICA charantia , *PEPTIDES , *TRYPSIN inhibitors , *MASS spectrometry , *NUCLEAR magnetic resonance spectroscopy , *NUCLEOTIDE sequence - Abstract
Two new peptides, MCh-1 and MCh-2, along with three known trypsin inhibitors (MCTI-I, MCTI-II and MCTI-III), were isolated from the seeds of the tropical vine Momordica charantia. The sequences of the peptides were determined using mass spectrometry and NMR spectroscopy. Using a strategy involving partial reduction and stepwise alkylation of the peptides, followed by enzymatic digestion and tandem mass spectrometry sequencing, the disulfide connectivity of MCh-1 was elucidated to be CysI-CysIV, CysII-CysV and CysIII-CysVI. The three-dimensional structures of MCh-1 and MCh-2 were determined using NMR spectroscopy and found to contain the inhibitor cystine knot (ICK) motif. The sequences of the novel peptides differ significantly from peptides previously isolated from this plant. Therefore, this study expands the known peptide diversity in M. charantia and the range of sequences that can be accommodated by the ICK motif. Furthermore, we show that a stable two-disulfide intermediate is involved in the oxidative folding of MCh-1. This disulfide intermediate is structurally homologous to the proposed ancestral fold of ICK peptides, and provides a possible pathway for the evolution of this structural motif, which is highly prevalent in nature. [ABSTRACT FROM AUTHOR]
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- 2013
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317. Isolation and characterization of α-conotoxin LsIA with potent activity at nicotinic acetylcholine receptors.
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Inserra, Marco C., Kompella, Shiva N., Vetter, Irina, Brust, Andreas, Daly, Norelle L., Cuny, Hartmut, Craik, David J., Alewood, Paul F., Adams, David J., and Lewis, Richard J.
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CONOTOXINS , *NICOTINIC acetylcholine receptors , *HIGH performance liquid chromatography , *NUCLEAR magnetic resonance spectroscopy , *PHARMACOLOGY , *N-terminal residues - Abstract
Abstract: A new α-conotoxin LsIA was isolated from the crude venom of Conus limpusi using assay-guided RP-HPLC fractionation. Synthetic LsIA was a potent antagonist of α3β2, α3α5β2 and α7 nAChRs, with half-maximal inhibitory concentrations of 10, 31 and 10nM, respectively. The structure of LsIA determined by NMR spectroscopy comprised a characteristic disulfide bond-stabilized α-helical structure and disordered N-terminal region. Potency reductions of up to 9-fold were observed for N-terminally truncated analogues of LsIA at α7 and α3β2 nAChRs, whereas C-terminal carboxylation enhanced potency 3-fold at α3β2 nAChRs but reduced potency 3-fold at α7 nAChRs. This study gives further insight into α-conotoxin pharmacology and the molecular basis of nAChR selectivity, highlighting the influence of N-terminal residues and C-terminal amidation on conotoxin pharmacology. [Copyright &y& Elsevier]
- Published
- 2013
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318. High-affinity Cyclic Peptide Matriptase Inhibitors.
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Quimbar, Pedro, Malik, Uru, Sommerhoff, Christian P., Kaas, Quentin, Chan, Lai Y., Huang, Yen-Hua, Grundhuber, Maresa, Dunse, Kerry, Craik, David J., Anderson, Marilyn A., and Daly, Norelle L.
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MATRIPTASE , *MEMBRANE proteins , *CELL proliferation , *SERINE proteinases , *STRUCTURE-activity relationships , *EXTRACELLULAR matrix - Abstract
The type II transmembrane serine protease matriptase is a key activator of multiple signaling pathways associated with cell proliferation and modification of the extracellular matrix. Deregulated matriptase activity correlates with a number of diseases, including cancer and hence highly selective matriptase inhibitors may have therapeutic potential. The plant-derived cyclic peptide, sunflower trypsin inhibitor-1 (SFTI-1), is a promising drug scaffold with potent matriptase inhibitory activity. In the current study we have analyzed the structure-activity relationships of SFTI-1 and Momordica cochinchinensis trypsin inhibitor-II (MCoTI-II), a structurally divergent trypsin inhibitor from Momordica cochinchinensis that also contains a cyclic backbone. We show that MCoTI-II is a significantly more potent matriptase inhibitor than SFTI-1 and that all alanine mutants of both peptides, generated using positional scanning mutagenesis, have decreased trypsin affinity, whereas several mutations either maintain or result in enhanced matriptase inhibitory activity. These intriguing results were used to design one of the most potent matriptase inhibitors known to date with a 290 pMequilibrium dissociation constant, and provide the first indication on how to modulate affinity for matriptase over trypsin in cyclic peptides. This information might be useful for the design of more selective and therapeutically relevant inhibitors of matriptase. [ABSTRACT FROM AUTHOR]
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- 2013
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319. A new family of cystine knot peptides from the seeds of Momordica cochinchinensis
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Chan, Lai Yue, He, Wenjun, Tan, Ninghua, Zeng, Guangzhi, Craik, David J., and Daly, Norelle L.
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CYSTINE , *PEPTIDES , *MOMORDICA , *TRYPSIN inhibitors , *ANTIMALARIALS , *PLASMODIUM falciparum , *ANTINEOPLASTIC agents - Abstract
Abstract: Momordica cochinchinensis, a Cucurbitaceae plant commonly found in Southeast Asia, has the unusual property of containing both acyclic and backbone-cyclized trypsin inhibitors with inhibitor cystine knot (ICK) motifs. In the current study we have shown that M. cochinchinensis also contains another family of acyclic ICK peptides. We recently reported two novel peptides from M. cochinchinensis but have now discovered four additional peptides (MCo-3–MCo-6) with related sequences. Together these peptides form a novel family of M. cochinchinensis ICK peptides (MCo-ICK) that do not have sequence homology with other known peptides and are not potent trypsin inhibitors. Otherwise these new peptides MCo-3 to MCo-6 were evaluated for antimalarial activity against Plasmodium falciparum, and cytotoxic activity against the cancer cell line MDA-MB-231. But these peptides were not active. [Copyright &y& Elsevier]
- Published
- 2013
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320. Cyclization of conotoxins to improve their biopharmaceutical properties
- Author
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Clark, Richard J., Akcan, Muharrem, Kaas, Quentin, Daly, Norelle L., and Craik, David J.
- Subjects
- *
CONOTOXINS , *RING formation (Chemistry) , *BIOPHARMACEUTICS , *CONUS , *PROTEOLYSIS , *DISULFIDES , *PEPTIDES , *ION channels , *DRUG design - Abstract
Abstract: Conotoxins are disulfide-rich peptides from the venoms of marine cone snails that are used in prey capture. Due to their exquisite potency and selectivity for different ion channels, receptors and transporters they have attracted much interest as leads in drug design. This article gives a brief background on conotoxins, describes their structures and highlights methods for synthetic cyclization to improve their biopharmaceutical properties. The proximity of the N and C termini of many conotoxins makes them particularly suitable for cyclization with linkers of on average five to seven amino acids. By linking the ends of conotoxins it is possible to significantly decrease their susceptibility to proteolysis without loss of their intrinsic biological activity. Here, the principles of conotoxin cyclization are illustrated with applications to the α- and χ- conotoxin classes, which have been implicated as leads for the treatment of pain and a range of other disorders including neuroprotection, schizophrenia, depression and cancer. [Copyright &y& Elsevier]
- Published
- 2012
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321. RegIIA: An α4/7-conotoxin from the venom of Conus regius that potently blocks α3β4 nAChRs
- Author
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Franco, Aldo, Kompella, Shiva N., Akondi, Kalyana B., Melaun, Christian, Daly, Norelle L., Luetje, Charles W., Alewood, Paul F., Craik, David J., Adams, David J., and Marí, Frank
- Subjects
- *
CONOTOXINS , *CHOLINERGIC receptors , *PERIPHERAL neuropathy , *CENTRAL nervous system abnormalities , *DRUG synergism , *ACETYLCHOLINE-binding proteins - Abstract
Abstract: Neuronal nicotinic acetylcholine receptors (nAChRs) play pivotal roles in the central and peripheral nervous systems. They are implicated in disease states such as Parkinson''s disease and schizophrenia, as well as addictive processes for nicotine and other drugs of abuse. Modulation of specific nAChRs is essential to understand their role in the CNS. α-Conotoxins, disulfide-constrained peptides isolated from the venom of cone snails, potently inhibit nAChRs. Their selectivity varies markedly depending upon the specific nAChR subtype/α-conotoxin pair under consideration. Thus, α-conotoxins are excellent probes to evaluate the functional roles of nAChRs subtypes. We isolated an α4/7-conotoxin (RegIIA) from the venom of Conus regius. Its sequence was determined by Edman degradation and confirmed by sequencing the cDNA of the protein precursor. RegIIA was synthesized using solid phase methods and native and synthetic RegIIA were functionally tested using two-electrode voltage clamp recording on nAChRs expressed in Xenopus laevis oocytes. RegIIA is among the most potent antagonist of the α3β4 nAChRs found to date and is also active at α3β2 and α7 nAChRs. The 3D structure of RegIIA reveals the typical folding of most α4/7-conotoxins. Thus, while structurally related to other α4/7 conotoxins, RegIIA has an exquisite balance of shape, charge, and polarity exposed in its structure to potently block the α3β4 nAChRs. [Copyright &y& Elsevier]
- Published
- 2012
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322. Isolation and characterization of cytotoxic cyclotides from Viola tricolor
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Tang, Jun, Wang, Conan K., Pan, Xulin, Yan, He, Zeng, Guangzhi, Xu, Wenyan, He, Wenjun, Daly, Norelle L., Craik, David J., and Tan, Ninghua
- Subjects
- *
PEPTIDE fractionation , *CELL-mediated cytotoxicity , *PLANT proteins , *WILD plants , *DRUG development , *NUCLEAR magnetic resonance spectroscopy , *MASS spectrometry - Abstract
Abstract: Many plants of the Violaceae plant family have been used in traditional remedies, and these plants often contain cyclotides, a particular type of plant cyclopeptide that is distinguished by a cyclic cystine knot motif. In general, bioactive plant cyclopeptides are interesting candidates for drug development. In the current study, a suite of 14 cyclotides, which includes seven novel cyclotides [vitri B, C, D, E, F, varv Hm, and He], together with seven known cyclotides [varv A, D, E, F, H, vitri A, and cycloviolacin O2], was isolated from Viola tricolor, a common flower. A chromatography-based method was used to isolate the cyclotides, which were characterized using tandem mass spectrometry and NMR spectroscopy. Several of the cyclotides showed cytotoxic activities against five cancer cell lines, U251, MDA-MB-231, A549, DU145, and BEL-7402. Three cyclotides, vitri A, vitri F, and cycloviolacin O2, were the most cytotoxic. The cytotoxic activity of the cyclotides did not correlate well with their hemolytic activity, indicating that different interactions, most likely with membranes, are involved for cytotoxic and hemolytic activities. Homology modeling of the structures was used in deriving structure–activity relationships. [Copyright &y& Elsevier]
- Published
- 2010
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323. Inhibition of Neuronal Nicotinic Acetyicholine Receptor Subtypes by α- Conotoxin GID and AnaIogues.
- Author
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MiIIard, Emma L., Nevin, Simon T., Loughnan, Marion L., Nick, Annette, Clark, Richard J., Alewood, Paul F., Lewis, Richard J., Adams, David J., Craik, David J., and Daly, Norelle L.
- Subjects
- *
TOXINS , *CONUS , *NICOTINIC receptors , *ACETYLCHOLINE , *BINDING sites , *CHRONIC pain treatment , *MAMMAL physiology , *THERAPEUTICS - Abstract
α-Conotoxins are small disulfide-rich peptides from the venom of the Conus species that target the nicotinic acetylcholine receptor (nAChR). They are valuable pharmacological tools and also have potential therapeutic applications particularly for the treatment of chronic pain. α-Conotoxin GID is isolated from the venom of Conus geographus and has an unusual N-terminal tail sequence that has been shown to be important for binding to the α4β2 subtype of the nAChR. To date, only four conotoxins that inhibit the α4β2 subtype have been characterized, but they are of considerable interest as it is the most abundant nAChR subtype in the mammalian brain and has been implicated in a range of diseases. In this study, analysis of alanine-scan and truncation mutants of GID reveals that a conserved proline in α-conotoxins is important for activity at the α7, α3β2, and α4β2 subtypes. Although the proline residue was the most critical residue for activity at the α3β2 subtype, Asp[sup3], Arg[sup12], and Asn[sup14] are also critical at the & subtype. Interestingly, very few of the mutations tested retained activity at the α4β2 subtype indicating a tightly defined binding site. This lack of tolerance to sequence variation may explain the lack of selective ligands discovered for the α4β2 subtype to date. Overall, our findings contribute to the understanding of the structureactivity relationships of α-conotoxins and may be beneficial for the ongoing attempts to exploit modulators of the neuronal nAChRs as therapeutic agents. [ABSTRACT FROM AUTHOR]
- Published
- 2009
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324. The A-chain of Human Relaxin Family Peptides Has Distinct Roles in the Binding and Activation of the Different Relaxin Family Peptide Receptors.
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Akhter Hossain, Mohammed, Rosengren, K. Johan, Haugaard-Jansson, Linda M., Soude Zhang, Layfield, Sharon, Ferraro, Tania, Daly, Norelle L., Tregear, Geoffrey W., Wade, John D., and Bathgate, Ross A. D.
- Subjects
- *
RELAXIN , *PEPTIDES , *NUCLEAR magnetic resonance spectroscopy , *ENZYME inhibitors , *HORMONES - Abstract
The relaxin peptides are a family of hormones that share a structural fold characterized by two chains, A and B, that are cross-braced by three disulfide bonds. Relaxins signal through two different classes of G-protein-coupled receptors (GPCRs), leucine-rich repeat-containing GPCRs LGR7 and LGR8 together with GPCR135 and GPCR142, now referred to as the relaxin family peptide (RXFP) receptors 1-4, respectively. Although key binding residues have been identified in the B-chain of the relaxin peptides, the role of the A-chain in their activity is currently unknown. A recent study showed that INSL3 can be truncated at the N terminus of its A-chain by up to 9 residues without affecting the binding affinity to its receptor RXFP2 while becoming a high affinity antagonist. This suggests that the N terminus of the INSL3 A-chain contains residues essential for RXFP2 activation. In this study, we have synthesized A-chain truncated human relaxin-2 and -3 (H2 and H3) relaxin peptides, characterized their structure by both CD and NMR spectroscopy, and tested their binding and cAMP activities on RXFP1, RXFP2, and RXFP3. In stark contrast to INSL3, A-chain-truncated H2 relaxin peptides lost RXFP1 and RXFP2 binding affinity and concurrently cAMP-stimulatory activity. H3 relaxin A-chain-truncated peptides displayed similar properties on RXFP1, highlighting a similar binding mechanism for H2 and H3 relaxin. In contrast, A-chain-truncated H3 relaxin peptides showed identical activity on RXFP3, highlighting that the B-chain is the sole determinant of the H3 relaxin-RXFP3 interaction. Our results provide new insights into the action of relaxins and demonstrate that the role of the A-chain for relaxin activity is both peptide- and receptor-dependent. [ABSTRACT FROM AUTHOR]
- Published
- 2008
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325. Isolation, Structure, and Activity of GID, A Novel alpha4/7-Conotoxin with an Externded N-terminal Sequence.
- Author
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Nicke, Annette, Loughnan, Marion L., Millar, Emma L., Alewwod, Paul F., Adams, David J., Daly, Norelle L., Craik, David J., and Lewis, Richard J.
- Subjects
- *
TOXINS , *DRUG receptors , *LIPIDS , *LIPOPROTEINS - Abstract
Studies the activity, structure and isolation of the novel alpha-4/7-conotoxin GID with an extended N-terminal sequence. Definition of the three-dimensional structure of GID over residues 4-19 with a similar motif to other alpha-conotoxins; Analysis of pertinent topics and relevant issues; Implications on lipids and lipoproteins.
- Published
- 2003
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326. Characterisation of predicted helical regions in the Chironex fleckeri CfTX-1 toxin.
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Andreosso, Athena, Bansal, Paramjit S., Smout, Michael J., Wilson, David, Seymour, Jamie E., and Daly, Norelle L.
- Subjects
- *
MARINE toxins , *JELLYFISHES , *ANTIVENINS , *PROTEIN structure ,CARDIOVASCULAR system pathophysiology - Published
- 2019
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327. Structural characterisation of defensive cone snail venom peptides.
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Wilson, David, Bansal, Paramjit, Dutertre, Sebastien, and Daly, Norelle L.
- Subjects
- *
VENOM , *CONUS , *PEPTIDES , *CONOTOXINS , *MEMBRANE proteins , *PHARMACOLOGY - Published
- 2019
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328. Solution structure of the N-terminal extension domain of a Schistosoma japonicum asparaginyl-tRNA synthetase.
- Author
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Peck Y, Pickering D, Mobli M, Liddell MJ, Wilson DT, Ruscher R, Ryan S, Buitrago G, McHugh C, Love NC, Pinlac T, Haertlein M, Kron MA, Loukas A, and Daly NL
- Subjects
- Animals, Models, Molecular, RNA, Transfer, Amino Acyl chemistry, RNA, Transfer, Amino Acyl metabolism, Helminth Proteins chemistry, Helminth Proteins metabolism, Protein Domains, Humans, Solutions, Schistosoma japonicum enzymology, Aspartate-tRNA Ligase chemistry, Aspartate-tRNA Ligase genetics, Aspartate-tRNA Ligase metabolism, Amino Acid Sequence
- Abstract
Several secreted proteins from helminths (parasitic worms) have been shown to have immunomodulatory activities. Asparaginyl-tRNA synthetases are abundantly secreted in the filarial nematode Brugia malayi ( Bm AsnRS) and the parasitic flatworm Schistosoma japonicum ( Sj AsnRS), indicating a possible immune function. The suggestion is supported by Bm AsnRS alleviating disease symptoms in a T-cell transfer mouse model of colitis. This immunomodulatory function is potentially related to an N-terminal extension domain present in eukaryotic AsnRS proteins but few structure/function studies have been done on this domain. Here we have determined the three-dimensional solution structure of the N-terminal extension domain of Sj AsnRS. A protein containing the 114 N-terminal amino acids of Sj AsnRS was recombinantly expressed with isotopic labelling to allow structure determination using 3D NMR spectroscopy, and analysis of dynamics using NMR relaxation experiments. Structural comparisons of the N-terminal extension domain of Sj AsnRS with filarial and human homologues highlight a high degree of variability in the β-hairpin region of these eukaryotic N-AsnRS proteins, but similarities in the disorder of the C-terminal regions. Limitations in PrDOS-based intrinsically disordered region (IDR) model predictions were also evident in this comparison. Empirical structural data such as that presented in our study for N- Sj AsnRS will enhance the prediction of sequence-homology based structure modelling and prediction of IDRs in the future.Communicated by Ramaswamy H. Sarma.
- Published
- 2024
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329. Author Correction: The NK cell granule protein NKG7 regulates cytotoxic granule exocytosis and inflammation.
- Author
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Ng SS, De Labastida Rivera F, Yan J, Corvino D, Das I, Zhang P, Kuns R, Chauhan SB, Hou J, Li XY, Frame TCM, McEnroe BA, Moore E, Na J, Engel JA, Soon MSF, Singh B, Kueh AJ, Herold MJ, Montes de Oca M, Singh SS, Bunn PT, Aguilera AR, Casey M, Braun M, Ghazanfari N, Wani S, Wang Y, Amante FH, Edwards CL, Haque A, Dougall WC, Singh OP, Baxter AG, Teng MWL, Loukas A, Daly NL, Cloonan N, Degli-Esposti MA, Uzonna J, Heath WR, Bald T, Tey SK, Nakamura K, Hill GR, Kumar R, Sundar S, Smyth MJ, and Engwerda CR
- Published
- 2024
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330. In silico design of potential Mcl-1 peptide-based inhibitors.
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Faraji N, Daly NL, Arab SS, and Khosroushahi AY
- Subjects
- Peptides pharmacology, Apoptosis, Myeloid Cell Leukemia Sequence 1 Protein chemistry, Myeloid Cell Leukemia Sequence 1 Protein metabolism, Cell Line, Tumor, bcl-X Protein, Apoptosis Regulatory Proteins chemistry, Apoptosis Regulatory Proteins genetics, Apoptosis Regulatory Proteins metabolism, Antineoplastic Agents pharmacology
- Abstract
Context: BIM (Bcl-2 interacting mediator of apoptosis)-derived peptides that specifically target over-expressed Mcl-1 (myeloid cell leukemia-1) protein and induce apoptosis are potentially anti-cancer agents. Since the helicity of BIM-derived peptides has a crucial role in their functionality, a range of strategies have been used to increase the helicity including the introduction of unnatural residues and stapling methods that have some drawbacks such as the accumulation in the liver. To avoid these drawbacks, this study aimed to design a more helical peptide by utilizing bioinformatics algorithms and molecular dynamics simulations without exploiting unnatural residues and stapling methods. MM-PBSA results showed that the mutations of A4fE and A2eE in analogue 5 demonstrate a preference towards binding with Mcl-1. As evidenced by Circular dichroism results, the helicity increases from 18 to 34%, these findings could enhance the potential of analogue 5 as an anti-cancer agent targeting Mcl-1. The applied strategies in this research could shed light on the in silico peptide design. Moreover, analogue 5 as a drug candidate can be evaluated in vitro and in vivo studies., Methods: The sequence of the lead peptide was determined using the ApInAPDB database and PRALINE program. Contact finder and PDBsum web server softwares were used to determine the contact involved amino acids in complex with Mcl-1. All identified salt bridge contributing residues were unaltered to preserve the binding affinity. After proposing novel analogues, their secondary structures were predicted by Cham finder web server software and GOR, Neural Network, and Chou-Fasman algorithms. Finally, molecular dynamics simulations run for 100 ns were done using the GROMACS, version 5.0.7, with the CHARMM36 force field. MM-PBSA was used to assess binding affinity specificity in targeting Mcl-1 and Bcl-xL (B-cell lymphoma extra-large)., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)
- Published
- 2024
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331. Structure-function and rational design of a spider toxin Ssp1a at human voltage-gated sodium channel subtypes.
- Author
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Dongol Y, Wilson DT, Daly NL, Cardoso FC, and Lewis RJ
- Abstract
The structure-function and optimization studies of Na
V -inhibiting spider toxins have focused on developing selective inhibitors for peripheral pain-sensing NaV 1.7. With several NaV subtypes emerging as potential therapeutic targets, structure-function analysis of NaV -inhibiting spider toxins at such subtypes is warranted. Using the recently discovered spider toxin Ssp1a, this study extends the structure-function relationships of NaV -inhibiting spider toxins beyond NaV 1.7 to include the epilepsy target NaV 1.2 and the pain target NaV 1.3. Based on these results and docking studies, we designed analogues for improved potency and/or subtype-selectivity, with S7R-E18K-rSsp1a and N14D-P27R-rSsp1a identified as promising leads. S7R-E18K-rSsp1a increased the rSsp1a potency at these three NaV subtypes, especially at NaV 1.3 (∼10-fold), while N14D-P27R-rSsp1a enhanced NaV 1.2/1.7 selectivity over NaV 1.3. This study highlights the challenge of developing subtype-selective spider toxin inhibitors across multiple NaV subtypes that might offer a more effective therapeutic approach. The findings of this study provide a basis for further rational design of Ssp1a and related NaSpTx1 homologs targeting NaV 1.2, NaV 1.3 and/or NaV 1.7 as research tools and therapeutic leads., 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. The authors declared that they were an editorial board member of Frontiers, at the time of submission. This had no impact on the peer review process and the final decision., (Copyright © 2023 Dongol, Wilson, Daly, Cardoso and Lewis.)- Published
- 2023
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332. Pmu1a, a novel spider toxin with dual inhibitory activity at pain targets hNa V 1.7 and hCa V 3 voltage-gated channels.
- Author
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Giribaldi J, Chemin J, Tuifua M, Deuis JR, Mary R, Vetter I, Wilson DT, Daly NL, Schroeder CI, Bourinet E, and Dutertre S
- Subjects
- Animals, Voltage-Gated Sodium Channel Blockers pharmacology, Voltage-Gated Sodium Channel Blockers chemistry, Pain, Peptides pharmacology, Magnetic Resonance Spectroscopy, Spider Venoms pharmacology, Spider Venoms chemistry, Spider Venoms metabolism, Spiders metabolism
- Abstract
Venom-derived peptides targeting ion channels involved in pain are regarded as a promising alternative to current, and often ineffective, chronic pain treatments. Many peptide toxins are known to specifically and potently block established therapeutic targets, among which the voltage-gated sodium and calcium channels are major contributors. Here, we report on the discovery and characterization of a novel spider toxin isolated from the crude venom of Pterinochilus murinus that shows inhibitory activity at both hNa
V 1.7 and hCaV 3.2 channels, two therapeutic targets implicated in pain pathways. Bioassay-guided HPLC fractionation revealed a 36-amino acid peptide with three disulfide bridges named μ/ω-theraphotoxin-Pmu1a (Pmu1a). Following isolation and characterization, the toxin was chemically synthesized and its biological activity was further assessed using electrophysiology, revealing Pmu1a to be a toxin that potently blocks both hNaV 1.7 and hCaV 3. Nuclear magnetic resonance structure determination of Pmu1a shows an inhibitor cystine knot fold that is the characteristic of many spider peptides. Combined, these data show the potential of Pmu1a as a basis for the design of compounds with dual activity at the therapeutically relevant hCaV 3.2 and hNaV 1.7 voltage-gated channels., (© 2023 Federation of European Biochemical Societies.)- Published
- 2023
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333. Peptides derived from hookworm anti-inflammatory proteins suppress inducible colitis in mice and inflammatory cytokine production by human cells.
- Author
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Cobos C, Bansal PS, Wilson DT, Jones L, Zhao G, Field MA, Eichenberger RM, Pickering DA, Ryan RYM, Ratnatunga CN, Miles JJ, Ruscher R, Giacomin PR, Navarro S, Loukas A, and Daly NL
- Abstract
A decline in the prevalence of parasites such as hookworms appears to be correlated with the rise in non-communicable inflammatory conditions in people from high- and middle-income countries. This correlation has led to studies that have identified proteins produced by hookworms that can suppress inflammatory bowel disease (IBD) and asthma in animal models. Hookworms secrete a family of abundant netrin-domain containing proteins referred to as AIPs (Anti-Inflammatory Proteins), but there is no information on the structure-function relationships. Here we have applied a downsizing approach to the hookworm AIPs to derive peptides of 20 residues or less, some of which display anti-inflammatory effects when co-cultured with human peripheral blood mononuclear cells and oral therapeutic activity in a chemically induced mouse model of acute colitis. Our results indicate that a conserved helical region is responsible, at least in part, for the anti-inflammatory effects. This helical region has potential in the design of improved leads for treating IBD and possibly other inflammatory conditions., 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 © 2022 Cobos, Bansal, Wilson, Jones, Zhao, Field, Eichenberger, Pickering, Ryan, Ratnatunga, Miles, Ruscher, Giacomin, Navarro, Loukas and Daly.)
- Published
- 2022
- Full Text
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334. Newly Discovered Peptides from the Coral Heliofungia actiniformis Show Structural and Functional Diversity.
- Author
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Schmidt CA, Cooke I, Wilson DT, Miller DJ, Peigneur S, Tytgat J, Field M, Takjoo R, Smout MJ, Loukas A, and Daly NL
- Subjects
- Animals, Biodiversity, Ecosystem, Humans, Peptides, Anthozoa
- Abstract
Scleractinian corals are crucially important to the health of some of the world's most biodiverse, productive, and economically important marine habitats. Despite this importance, analysis of coral peptidomes is still in its infancy. Here we show that the tentacle extract from the stony coral Heliofungia actiniformis is rich in peptides with diverse and novel structures. We have characterized the sequences and three-dimensional structures of four new peptides, three of which have no known homologues. We show that a 2 kDa peptide, Hact-2, promotes significant cell proliferation on human cells and speculate this peptide may be involved in the remarkable regenerative capacity of corals. We found a 3 kDa peptide, Hact-3, encoded within a fascin-like domain, and homologues of Hact-3 are present in the genomes of other coral species. Two additional peptides, Hact-4 and Hact-SCRiP1, with limited sequence similarity, both contain a beta-defensin-like fold and highlight a structural link with the small cysteine-rich proteins (SCRiP) family of proteins found predominantly in corals. Our results provide a first glimpse into the remarkable and unexplored structural diversity of coral peptides, providing insight into their diversity and putative functions and, given the ancient lineage of corals, potential insight into the evolution of structural motifs.
- Published
- 2022
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335. Voltage-Gated Sodium Channel Modulation by a New Spider Toxin Ssp1a Isolated From an Australian Theraphosid.
- Author
-
Dongol Y, Choi PM, Wilson DT, Daly NL, Cardoso FC, and Lewis RJ
- Abstract
Given the important role of voltage-gated sodium (Na
V ) channel-modulating spider toxins in elucidating the function, pharmacology, and mechanism of action of therapeutically relevant NaV channels, we screened the venom from Australian theraphosid species against the human pain target hNaV 1.7. Using assay-guided fractionation, we isolated a 33-residue inhibitor cystine knot (ICK) peptide (Ssp1a) belonging to the NaSpTx1 family. Recombinant Ssp1a (rSsp1a) inhibited neuronal hNaV subtypes with a rank order of potency hNaV 1.7 > 1.6 > 1.2 > 1.3 > 1.1. rSsp1a inhibited hNaV 1.7, hNaV 1.2 and hNaV 1.3 without significantly altering the voltage-dependence of activation, inactivation, or delay in recovery from inactivation. However, rSsp1a demonstrated voltage-dependent inhibition at hNaV 1.7 and rSsp1a-bound hNaV 1.7 opened at extreme depolarizations, suggesting rSsp1a likely interacted with voltage-sensing domain II (VSD II) of hNaV 1.7 to trap the channel in its resting state. Nuclear magnetic resonance spectroscopy revealed key structural features of Ssp1a, including an amphipathic surface with hydrophobic and charged patches shown by docking studies to comprise the interacting surface. This study provides the basis for future structure-function studies to guide the development of subtype selective inhibitors., 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 Dongol, Choi, Wilson, Daly, Cardoso and Lewis.)- Published
- 2021
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336. Plant derived cyclic peptides.
- Author
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Daly NL and Wilson DT
- Subjects
- Animals, Cyclotides chemistry, Cyclotides pharmacology, Cysteine Endopeptidases metabolism, Disease Models, Animal, Encephalomyelitis, Autoimmune, Experimental drug therapy, Humans, Multiple Sclerosis drug therapy, Peptides, Cyclic chemistry, Peptides, Cyclic pharmacology, Plant Proteins chemistry, Plant Proteins pharmacology, Protein Precursors chemistry, Protein Precursors metabolism, Cyclotides metabolism, Peptides, Cyclic metabolism, Plant Proteins metabolism, Plants metabolism
- Abstract
Cyclic peptides are widespread throughout the plant kingdom, and display diverse sequences, structures and bioactivities. The potential applications attributed to these peptides and their unusual biosynthesis has captivated the attention of researchers for many years. Several gene sequences for plant cyclic peptides have been discovered over the last two decades but it is only recently that we are beginning to understand the intricacies associated with their biosynthesis. Recent studies have focussed on three main classes of plant derived cyclic peptides, namely orbitides, SFTI related peptides and cyclotides. In this mini-review, we discuss the expansion of the known sequence and structural diversity in these families, insights into the enzymes involved in the biosynthesis, the exciting applications which includes a cyclotide currently in clinical trials for the treatment of multiple sclerosis, and new production methods that are being developed to realise the potential of plant cyclic peptides as pharmaceutical or agricultural agents., (© 2021 The Author(s).)
- Published
- 2021
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337. A netrin domain-containing protein secreted by the human hookworm Necator americanus protects against CD4 T cell transfer colitis.
- Author
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Buitrago G, Pickering D, Ruscher R, Cobos Caceres C, Jones L, Cooper M, Van Waardenberg A, Ryan S, Miles K, Field M, Dredge K, Daly NL, Giacomin PR, and Loukas A
- Subjects
- Animals, CD4-Positive T-Lymphocytes transplantation, Colitis, Ulcerative drug therapy, Colitis, Ulcerative immunology, Disease Models, Animal, Female, Helminth Proteins chemistry, Helminth Proteins genetics, Hookworm Infections metabolism, Humans, Male, Matrix Metalloproteinase Inhibitors chemistry, Mice, Inbred C57BL, Mice, Knockout, Netrins analysis, Recombinant Proteins administration & dosage, Mice, Anti-Inflammatory Agents administration & dosage, CD4-Positive T-Lymphocytes immunology, Colitis, Ulcerative prevention & control, Helminth Proteins administration & dosage, Necator americanus chemistry, Netrins administration & dosage
- Abstract
The symbiotic relationships shared between humans and their gastrointestinal parasites present opportunities to discover novel therapies for inflammatory diseases. A prime example of this phenomenon is the interaction of humans and roundworms such as the hookworm, Necator americanus. Epidemiological observations, animal studies and clinical trials using experimental human hookworm infection show that hookworms can suppress inflammation in a safe and well-tolerated way, and that the key to their immunomodulatory properties lies within their secreted proteome. Herein we describe the identification of 2 netrin domain-containing proteins from the N. americanus secretome, and explore their potential in treating intestinal inflammation in mouse models of ulcerative colitis. One of these proteins, subsequently named Na-AIP-1, was effective at suppressing disease when administered prophylactically in the acute TNBS-induced model of colitis. This protective effect was validated in the more robust CD4 T cell transfer model of chronic colitis, where prophylactic Na-AIP-1 reduced T-cell-dependent type-1 cytokine responses in the intestine and the associated intestinal pathology. Mechanistic studies revealed that depletion of CD11c+ cells abrogated the protective anticolitic effect of Na-AIP-1. Next generation sequencing of colon tissue in the T-cell transfer model of colitis revealed that Na-AIP-1 induced a transcriptomic profile associated with the downregulation of metabolic and signaling pathways involved in type-1 inflammation, notably TNF. Finally, co-culture of Na-AIP-1 with a human monocyte-derived M1 macrophage cell line resulted in significantly reduced secretion of TNF. Na-AIP-1 is now a candidate for clinical development as a novel therapeutic for the treatment of human inflammatory bowel diseases., (Copyright © 2021 The Author(s). Published by Elsevier Inc. All rights reserved.)
- Published
- 2021
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338. Gastrointestinal Helminth Infection Improves Insulin Sensitivity, Decreases Systemic Inflammation, and Alters the Composition of Gut Microbiota in Distinct Mouse Models of Type 2 Diabetes.
- Author
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Khudhair Z, Alhallaf R, Eichenberger RM, Whan J, Kupz A, Field M, Krause L, Wilson DT, Daly NL, Giacomin P, Sotillo J, and Loukas A
- Subjects
- Animals, Blood Glucose, Body Weight, Diabetes Mellitus, Type 2 etiology, Diabetes Mellitus, Type 2 metabolism, Diet, High-Fat, Eosinophils, Glucose Tolerance Test, Leukocyte Count, Male, Metabolic Syndrome therapy, Mice, Mice, Inbred C57BL, Diabetes Mellitus, Type 2 therapy, Gastrointestinal Microbiome, Inflammation prevention & control, Insulin Resistance, Nippostrongylus, Strongylida Infections physiopathology
- Abstract
Type 2 diabetes (T2D) is a major health problem and is considered one of the top 10 diseases leading to death globally. T2D has been widely associated with systemic and local inflammatory responses and with alterations in the gut microbiota. Microorganisms, including parasitic worms and gut microbes have exquisitely co-evolved with their hosts to establish an immunological interaction that is essential for the formation and maintenance of a balanced immune system, including suppression of excessive inflammation. Herein we show that both prophylactic and therapeutic infection of mice with the parasitic hookworm-like nematode, Nippostrongylus brasiliensis , significantly reduced fasting blood glucose, oral glucose tolerance and body weight gain in two different diet-induced mouse models of T2D. Helminth infection was associated with elevated type 2 immune responses including increased eosinophil numbers in the mesenteric lymph nodes, liver and adipose tissues, as well as increased expression of IL-4 and alternatively activated macrophage marker genes in adipose tissue, liver and gut. N. brasiliensis infection was also associated with significant compositional changes in the gut microbiota at both the phylum and order levels. Our findings show that N. brasiliensis infection drives changes in local and systemic immune cell populations, and that these changes are associated with a reduction in systemic and local inflammation and compositional changes in the gut microbiota which cumulatively might be responsible for the improved insulin sensitivity observed in infected mice. Our findings indicate that carefully controlled therapeutic hookworm infection in humans could be a novel approach for treating metabolic syndrome and thereby preventing T2D., Competing Interests: Author LK was employed by company Microba Pty Ltd. The remaining 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 Khudhair, Alhallaf, Eichenberger, Whan, Kupz, Field, Krause, Wilson, Daly, Giacomin, Sotillo and Loukas.)
- Published
- 2021
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339. ampir: an R package for fast genome-wide prediction of antimicrobial peptides.
- Author
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Fingerhut LCHW, Miller DJ, Strugnell JM, Daly NL, and Cooke IR
- Subjects
- Machine Learning, Pore Forming Cytotoxic Proteins, Genome, Software
- Abstract
Summary: Antimicrobial peptides (AMPs) are the key components of the innate immune system that protect against pathogens, regulate the microbiome and are promising targets for pharmaceutical research. Computational tools based on machine learning have the potential to aid discovery of genes encoding novel AMPs but existing approaches are not designed for genome-wide scans. To facilitate such genome-wide discovery of AMPs we developed a fast and accurate AMP classification framework, ampir. ampir is designed for high throughput, integrates well with existing bioinformatics pipelines, and has much higher classification accuracy than existing methods when applied to whole genome data., Availability and Implementation: ampir is implemented primarily in R with core feature calculation methods written in C++. Release versions are available via CRAN and work on all major operating systems. The development version is maintained at https://github.com/legana/ampir., Supplementary Information: Supplementary data are available at Bioinformatics online., (© The Author(s) 2020. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)
- Published
- 2021
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340. Identification and Characterization of a Peptide from the Stony Coral Heliofungia actiniformis .
- Author
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Schmidt CA, Wilson DT, Cooke I, Potriquet J, Tungatt K, Muruganandah V, Boote C, Kuek F, Miles JJ, Kupz A, Ryan S, Loukas A, Bansal PS, Takjoo R, Miller DJ, Peigneur S, Tytgat J, and Daly NL
- Subjects
- Amino Acid Sequence, Animals, Anti-Bacterial Agents pharmacology, Chromatography, High Pressure Liquid methods, Gram-Negative Bacteria drug effects, Gram-Positive Bacteria drug effects, Magnetic Resonance Spectroscopy methods, Mass Spectrometry methods, Peptides pharmacology, Anthozoa chemistry, Anti-Bacterial Agents chemistry, Peptides chemistry
- Abstract
Marine organisms produce a diverse range of toxins and bioactive peptides to support predation, competition, and defense. The peptide repertoires of stony corals (order Scleractinia) remain relatively understudied despite the presence of tentacles used for predation and defense that are likely to contain a range of bioactive compounds. Here, we show that a tentacle extract from the mushroom coral, Heliofungia actiniformis , contains numerous peptides with a range of molecular weights analogous to venom profiles from species such as cone snails. Using NMR spectroscopy and mass spectrometry we characterized a 12-residue peptide (Hact-1) with a new sequence (GCHYTPFGLICF) and well-defined β-hairpin structure stabilized by a single disulfide bond. The sequence is encoded within the genome of the coral and expressed in the polyp body tissue. The structure present is common among toxins and venom peptides, but Hact-1 does not show activity against select examples of Gram-positive and Gram-negative bacteria or a range of ion channels, common properties of such peptides. Instead, it appears to have a limited effect on human peripheral blood mononuclear cells, but the ecological function of the peptide remains unknown. The discovery of this peptide from H. actiniformis is likely to be the first of many from this and related species.
- Published
- 2020
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341. Backbone Cyclization Turns a Venom Peptide into a Stable and Equipotent Ligand at Both Muscle and Neuronal Nicotinic Receptors.
- Author
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Giribaldi J, Haufe Y, Evans ERJ, Amar M, Durner A, Schmidt C, Faucherre A, Moha Ou Maati H, Enjalbal C, Molgó J, Servent D, Wilson DT, Daly NL, Nicke A, and Dutertre S
- Subjects
- Amino Acid Sequence, Animals, Conotoxins chemistry, Cyclization, Larva drug effects, Larva physiology, Locomotion drug effects, Mice, Muscle Contraction drug effects, Nicotinic Antagonists metabolism, Nicotinic Antagonists pharmacology, Peptides metabolism, Peptides pharmacology, Protein Binding, Protein Structure, Tertiary, Receptors, Nicotinic chemistry, Zebrafish growth & development, Zebrafish physiology, Ligands, Muscles metabolism, Neurons metabolism, Nicotinic Antagonists chemistry, Peptides chemistry, Receptors, Nicotinic metabolism, Venoms metabolism
- Abstract
Venom peptides are promising drug leads, but their therapeutic use is often limited by stability and bioavailability issues. In this study, we designed cyclic analogues of α-conotoxin CIA, a potent muscle nicotinic acetylcholine receptor (nAChR) blocker with a significantly lower affinity at the neuronal α3β2 subtype. Remarkably, all analogues retained the low nanomolar activity of native CIA toward muscle-type nAChRs but showed greatly improved resistance to degradation in human serum and, surprisingly, displayed up to 52-fold higher potency for the α3β2 neuronal nAChR subtype (IC
50 1.3 nM). Comparison of nuclear magnetic resonance-derived structures revealed some differences that might explain the gain of potency at α3β2 nAChRs. All peptides were highly paralytic when injected into adult zebrafish and bath-applied to zebrafish larvae, suggesting barrier-crossing capabilities and efficient uptake. Finally, these cyclic CIA analogues were shown to be unique pharmacological tools to investigate the contribution of the presynaptic α3β2 nAChR subtype to the train-of-four fade.- Published
- 2020
- Full Text
- View/download PDF
342. The NK cell granule protein NKG7 regulates cytotoxic granule exocytosis and inflammation.
- Author
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Ng SS, De Labastida Rivera F, Yan J, Corvino D, Das I, Zhang P, Kuns R, Chauhan SB, Hou J, Li XY, Frame TCM, McEnroe BA, Moore E, Na J, Engel JA, Soon MSF, Singh B, Kueh AJ, Herold MJ, Montes de Oca M, Singh SS, Bunn PT, Aguilera AR, Casey M, Braun M, Ghazanfari N, Wani S, Wang Y, Amante FH, Edwards CL, Haque A, Dougall WC, Singh OP, Baxter AG, Teng MWL, Loukas A, Daly NL, Cloonan N, Degli-Esposti MA, Uzonna J, Heath WR, Bald T, Tey SK, Nakamura K, Hill GR, Kumar R, Sundar S, Smyth MJ, and Engwerda CR
- Subjects
- Animals, Cells, Cultured, Cytotoxicity, Immunologic, Disease Models, Animal, Exocytosis, Humans, Membrane Proteins genetics, Mice, Mice, Inbred C57BL, Secretory Vesicles metabolism, CD4-Positive T-Lymphocytes immunology, CD8-Positive T-Lymphocytes immunology, Inflammation immunology, Killer Cells, Natural immunology, Leishmania donovani physiology, Leishmaniasis, Visceral immunology, Malaria immunology, Membrane Proteins metabolism, Plasmodium physiology
- Abstract
Immune-modulating therapies have revolutionized the treatment of chronic diseases, particularly cancer. However, their success is restricted and there is a need to identify new therapeutic targets. Here, we show that natural killer cell granule protein 7 (NKG7) is a regulator of lymphocyte granule exocytosis and downstream inflammation in a broad range of diseases. NKG7 expressed by CD4
+ and CD8+ T cells played key roles in promoting inflammation during visceral leishmaniasis and malaria-two important parasitic diseases. Additionally, NKG7 expressed by natural killer cells was critical for controlling cancer initiation, growth and metastasis. NKG7 function in natural killer and CD8+ T cells was linked with their ability to regulate the translocation of CD107a to the cell surface and kill cellular targets, while NKG7 also had a major impact on CD4+ T cell activation following infection. Thus, we report a novel therapeutic target expressed on a range of immune cells with functions in different immune responses.- Published
- 2020
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343. A C-Terminal Fragment of Chlorotoxin Retains Bioactivity and Inhibits Cell Migration.
- Author
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Dastpeyman M, Giacomin P, Wilson D, Nolan MJ, Bansal PS, and Daly NL
- Abstract
Chlorotoxin was originally isolated from the venom of the Israeli scorpion Leiurus quinquestriatus , and has potential as a tumor imaging agent based on its selective binding to tumor cells. Several targets have been suggested for chlorotoxin including voltage-gated chloride channels, and it has been shown to have anti-angiogenic activity and inhibit cell migration. The structure of chlorotoxin is stabilized by four disulfide bonds and contains β-sheet and helical structure. Interestingly, the reduced form has previously been shown to inhibit cell migration to the same extent as the wild type, but structural analysis indicates that the reduced form of the peptide does not maintain the native secondary structure and appears unstructured in solution. This lack of structure suggests that a short stretch of amino acids might be responsible for the bioactivity. To explore this hypothesis, we have synthesized fragments of chlorotoxin without disulfide bonds. As expected for such small peptides, NMR analysis indicated that the peptides were unstructured in solution. However, the peptide corresponding to the eight C-terminal residues inhibited cell migration, in contrast to the other fragments. Our results suggest that the C-terminal region plays a critical role in the bioactivity of chlorotoxin.
- Published
- 2019
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344. Structural Variants of a Liver Fluke Derived Granulin Peptide Potently Stimulate Wound Healing.
- Author
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Dastpeyman M, Bansal PS, Wilson D, Sotillo J, Brindley PJ, Loukas A, Smout MJ, and Daly NL
- Subjects
- Animals, Fasciola hepatica metabolism, Fascioliasis parasitology, Female, Granulins chemistry, Helminth Proteins chemistry, Mice, Mice, Inbred BALB C, Peptide Fragments chemistry, Protein Conformation, Cell Proliferation drug effects, Fasciola hepatica chemistry, Granulins pharmacology, Helminth Proteins pharmacology, Peptide Fragments pharmacology, Skin Diseases prevention & control, Wound Healing drug effects
- Abstract
Granulins are a family of growth factors involved in cell proliferation. The liver-fluke granulin, Ov-GRN-1, isolated from a carcinogenic liver fluke Opisthorchis viverrini, can significantly accelerate wound repair in vivo and in vitro. However, it is difficult to express Ov-GRN-1 in recombinant form at high yield, impeding its utility as a drug lead. Previously we reported that a truncated analogue ( Ov-GRN
12-35_3s ) promotes healing of cutaneous wounds in mice. NMR analysis of this analogue indicates the presence of multiple conformations, most likely as a result of proline cis/ trans isomerization. To further investigate whether the proline residues are involved in adopting the multiple confirmations, we have synthesized analogues involving mutation of the proline residues. We have shown that the proline residues have a significant influence on the structure, activity, and folding of Ov-GRN12-35_3s . These results provide insight into improving the oxidative folding yield and bioactivity of Ov-GRN12-35_3s and might facilitate the development of a novel wound healing agent.- Published
- 2018
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- View/download PDF
345. Structural diversity of arthropod venom toxins.
- Author
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Daly NL and Wilson D
- Subjects
- Amino Acid Motifs, Animals, Arthropods, Structure-Activity Relationship, Arthropod Venoms chemistry, Peptides chemistry, Proteins chemistry
- Abstract
Arthropods are a diverse and ancient group of invertebrate animals, which constitute approximately 75-85% of all known species on earth. Many arthropod species, such as spiders, scorpions and even some crustaceans, contain venoms that can be very complex, representing natural combinatorial libraries of bioactive compounds. Characterization of the compounds in these libraries has helped the development of tools for pharmacology, and the discovery of lead molecules for therapeutic treatments. A critical aspect in the characterization of venom compounds is determination of three-dimensional structure. Structural analysis can provide insight into many processes including understanding target interactions and developing more potent and selective drug leads. Arthropod venoms are an extremely rich source of novel structures and this review provides an overview of this structural diversity, including structures of proteins, peptides and small molecules., (Copyright © 2018 Elsevier Ltd. All rights reserved.)
- Published
- 2018
- Full Text
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346. Venomics: A Mini-Review.
- Author
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Wilson D and Daly NL
- Abstract
Venomics is the integration of proteomic, genomic and transcriptomic approaches to study venoms. Advances in these approaches have enabled increasingly more comprehensive analyses of venoms to be carried out, overcoming to some extent the limitations imposed by the complexity of the venoms and the small quantities that are often available. Advances in bioinformatics and high-throughput functional assay screening approaches have also had a significant impact on venomics. A combination of all these techniques is critical for enhancing our knowledge on the complexity of venoms and their potential therapeutic and agricultural applications. Here we highlight recent advances in these fields and their impact on venom analyses.
- Published
- 2018
- Full Text
- View/download PDF
347. Corrigendum: Vicinal Disulfide Constrained Cyclic Peptidomimetics: a Turn Mimetic Scaffold Targeting the Norepinephrine Transporter.
- Author
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Brust A, Wang CA, Daly NL, Kennerly J, Sadeghi M, Christie MJ, Lewis RJ, Mobli M, and Alewood PF
- Published
- 2018
- Full Text
- View/download PDF
348. Structure and Biological Activity of a Turripeptide from Unedogemmula bisaya Venom.
- Author
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Omaga CA, Carpio LD, Imperial JS, Daly NL, Gajewiak J, Flores MS, Espino SS, Christensen S, Filchakova OM, López-Vera E, Raghuraman S, Olivera BM, and Concepcion GP
- Subjects
- Animals, Calcium metabolism, Cells, Cultured, Conotoxins isolation & purification, Conus Snail drug effects, Conus Snail genetics, Conus Snail growth & development, Female, Ganglia, Spinal cytology, Ganglia, Spinal drug effects, Ganglia, Spinal metabolism, Male, Mice, Mice, Inbred ICR, Models, Molecular, Neurons cytology, Neurons drug effects, Neurons metabolism, Oocytes cytology, Oocytes drug effects, Oocytes metabolism, Peptide Fragments chemistry, Peptide Fragments pharmacology, Receptors, Nicotinic metabolism, Xenopus laevis, Conotoxins chemistry, Conotoxins pharmacology, Conus Snail chemistry
- Abstract
The turripeptide ubi3a was isolated from the venom of the marine gastropod Unedogemmula bisaya, family Turridae, by bioassay-guided purification; both native and synthetic ubi3a elicited prolonged tremors when injected intracranially into mice. The sequence of the peptide, DCCOCOAGAVRCRFACC-NH
2 (O = 4-hydroxyproline) follows the framework III pattern for cysteines (CC-C-C-CC) in the M-superfamily of conopeptides. The three-dimensional structure determined by NMR spectroscopy indicated a disulfide connectivity that is not found in conopeptides with the cysteine framework III: C1 -C4, C2 -C6 , C3 -C5 . The peptide inhibited the activity of the α9α10 nicotinic acetylcholine receptor with relatively low affinity (IC50 , 10.2 μM). Initial Constellation Pharmacology data revealed an excitatory activity of ubi3a on a specific subset of mouse dorsal root ganglion neurons.- Published
- 2017
- Full Text
- View/download PDF
349. An engineered cyclic peptide alleviates symptoms of inflammation in a murine model of inflammatory bowel disease.
- Author
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Cobos Caceres C, Bansal PS, Navarro S, Wilson D, Don L, Giacomin P, Loukas A, and Daly NL
- Subjects
- Animals, Anti-Inflammatory Agents, Non-Steroidal chemistry, Anti-Inflammatory Agents, Non-Steroidal metabolism, Colitis, Ulcerative immunology, Colitis, Ulcerative pathology, Colon immunology, Colon pathology, Drug Design, Drug Stability, Gastrointestinal Agents chemical synthesis, Gastrointestinal Agents chemistry, Humans, Male, Mice, Inbred C57BL, Organ Size drug effects, Peptides, Cyclic chemistry, Peptides, Cyclic metabolism, Protein Conformation, Protein Engineering, Protein Folding, Protein Stability, Proteolysis, Random Allocation, Serum enzymology, Specific Pathogen-Free Organisms, Anti-Inflammatory Agents, Non-Steroidal therapeutic use, Colitis, Ulcerative drug therapy, Colon drug effects, Disease Models, Animal, Gastrointestinal Agents therapeutic use, Models, Molecular, Peptides, Cyclic therapeutic use
- Abstract
Inflammatory bowel diseases (IBDs) are a set of complex and debilitating diseases for which there is no satisfactory treatment. Recent studies have shown that small peptides show promise for reducing inflammation in models of IBD. However, these small peptides are likely to be unstable and rapidly cleared from the circulation, and therefore, if not modified for better stability, represent non-viable drug leads. We hypothesized that improving the stability of these peptides by grafting them into a stable cyclic peptide scaffold may enhance their therapeutic potential. Using this approach, we have designed a novel cyclic peptide that comprises a small bioactive peptide from the annexin A1 protein grafted into a sunflower trypsin inhibitor cyclic scaffold. We used native chemical ligation to synthesize the grafted cyclic peptide. This engineered cyclic peptide maintained the overall fold of the naturally occurring cyclic peptide, was more effective at reducing inflammation in a mouse model of acute colitis than the bioactive peptide alone, and showed enhanced stability in human serum. Our findings suggest that the use of cyclic peptides as structural backbones offers a promising approach for the treatment of IBD and potentially other chronic inflammatory conditions., Competing Interests: The authors declare that they have no conflicts of interest with the contents of this article., (© 2017 by The American Society for Biochemistry and Molecular Biology, Inc.)
- Published
- 2017
- Full Text
- View/download PDF
350. The N-terminal pro-domain of the kalata B1 cyclotide precursor is intrinsically unstructured.
- Author
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Daly NL, Gunasekera S, Clark RJ, Lin F, Wade JD, Anderson MA, and Craik DJ
- Subjects
- Protein Domains, Cyclotides chemistry, Intrinsically Disordered Proteins chemistry, Oldenlandia chemistry, Plant Proteins chemistry
- Abstract
Cyclotides are plant-derived, gene-encoded, circular peptides with a range of host-defense functions, including insecticidal activity. They also have potential as pharmaceutical scaffolds and understanding their biosynthesis is important to facilitate their large-scale production. Insights into the biosynthesis of cyclotides are emerging but there are still open questions, particularly regarding the influence of the structure of the precursor proteins on processing/biosynthetic pathways. The precursor protein of kalata B1, encoded by the plant Oldenlandia affinis, contains N- and C-terminal propeptides that flank the mature cyclotide domain. The C-terminal region (ctr) is important for the cyclization process, whereas the N-terminal repeat (ntr) has been implicated in vacuolar targeting. In this study we examined the structure and folding of various truncated constructs of the ntr coupled to the mature domain of kalata B1. Despite the ntr having a well-defined helical structure in isolation, once coupled to the natively folded mature domain there is no evidence of an ordered structure. Surprisingly, the ntr appears to be highly disordered and induces self-association of the precursor. This self-association might be associated with the role of the ntr as a vacuolar-targeting signal, as previously shown for unrelated storage proteins., (© 2016 Wiley Periodicals, Inc.)
- Published
- 2016
- Full Text
- View/download PDF
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