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Ero1-Mediated Reoxidation of Protein Disulfide Isomerase Accelerates the Folding of Cone Snail Toxins

Disulfide-rich peptides are highly abundant in nature and their study has provided fascinating insight into protein folding, structure and function. Venomous cone snails belong to a group of organisms that express one of the largest sets of disulfide-rich peptides (conotoxins) found in nature. The d...

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Autores principales: O’Brien, Henrik, Kanemura, Shingo, Okumura, Masaki, Baskin, Robert P., Bandyopadhyay, Pradip K., Olivera, Baldomero M., Ellgaard, Lars, Inaba, Kenji, Safavi-Hemami, Helena
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6275033/
https://www.ncbi.nlm.nih.gov/pubmed/30384459
http://dx.doi.org/10.3390/ijms19113418
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author O’Brien, Henrik
Kanemura, Shingo
Okumura, Masaki
Baskin, Robert P.
Bandyopadhyay, Pradip K.
Olivera, Baldomero M.
Ellgaard, Lars
Inaba, Kenji
Safavi-Hemami, Helena
author_facet O’Brien, Henrik
Kanemura, Shingo
Okumura, Masaki
Baskin, Robert P.
Bandyopadhyay, Pradip K.
Olivera, Baldomero M.
Ellgaard, Lars
Inaba, Kenji
Safavi-Hemami, Helena
author_sort O’Brien, Henrik
collection PubMed
description Disulfide-rich peptides are highly abundant in nature and their study has provided fascinating insight into protein folding, structure and function. Venomous cone snails belong to a group of organisms that express one of the largest sets of disulfide-rich peptides (conotoxins) found in nature. The diversity of structural scaffolds found for conotoxins suggests that specialized molecular adaptations have evolved to ensure their efficient folding and secretion. We recently showed that canonical protein disulfide isomerase (PDI) and a conotoxin-specific PDI (csPDI) are ubiquitously expressed in the venom gland of cone snails and play a major role in conotoxin folding. Here, we identify cone snail endoplasmic reticulum oxidoreductin-1 (Conus Ero1) and investigate its role in the oxidative folding of conotoxins through reoxidation of cone snail PDI and csPDI. We show that Conus Ero1 preferentially reoxidizes PDI over csPDI, suggesting that the reoxidation of csPDI may rely on an Ero1-independent molecular pathway. Despite the preferential reoxidation of PDI over csPDI, the combinatorial effect of Ero1 and csPDI provides higher folding yields than Ero1 and PDI. We further demonstrate that the highest in vitro folding rates of two model conotoxins are achieved when all three enzymes are present, indicating that these enzymes may act synergistically. Our findings provide new insight into the generation of one of the most diverse classes of disulfide-rich peptides and may improve current in vitro approaches for the production of venom peptides for pharmacological studies.
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spelling pubmed-62750332018-12-15 Ero1-Mediated Reoxidation of Protein Disulfide Isomerase Accelerates the Folding of Cone Snail Toxins O’Brien, Henrik Kanemura, Shingo Okumura, Masaki Baskin, Robert P. Bandyopadhyay, Pradip K. Olivera, Baldomero M. Ellgaard, Lars Inaba, Kenji Safavi-Hemami, Helena Int J Mol Sci Article Disulfide-rich peptides are highly abundant in nature and their study has provided fascinating insight into protein folding, structure and function. Venomous cone snails belong to a group of organisms that express one of the largest sets of disulfide-rich peptides (conotoxins) found in nature. The diversity of structural scaffolds found for conotoxins suggests that specialized molecular adaptations have evolved to ensure their efficient folding and secretion. We recently showed that canonical protein disulfide isomerase (PDI) and a conotoxin-specific PDI (csPDI) are ubiquitously expressed in the venom gland of cone snails and play a major role in conotoxin folding. Here, we identify cone snail endoplasmic reticulum oxidoreductin-1 (Conus Ero1) and investigate its role in the oxidative folding of conotoxins through reoxidation of cone snail PDI and csPDI. We show that Conus Ero1 preferentially reoxidizes PDI over csPDI, suggesting that the reoxidation of csPDI may rely on an Ero1-independent molecular pathway. Despite the preferential reoxidation of PDI over csPDI, the combinatorial effect of Ero1 and csPDI provides higher folding yields than Ero1 and PDI. We further demonstrate that the highest in vitro folding rates of two model conotoxins are achieved when all three enzymes are present, indicating that these enzymes may act synergistically. Our findings provide new insight into the generation of one of the most diverse classes of disulfide-rich peptides and may improve current in vitro approaches for the production of venom peptides for pharmacological studies. MDPI 2018-10-31 /pmc/articles/PMC6275033/ /pubmed/30384459 http://dx.doi.org/10.3390/ijms19113418 Text en © 2018 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
O’Brien, Henrik
Kanemura, Shingo
Okumura, Masaki
Baskin, Robert P.
Bandyopadhyay, Pradip K.
Olivera, Baldomero M.
Ellgaard, Lars
Inaba, Kenji
Safavi-Hemami, Helena
Ero1-Mediated Reoxidation of Protein Disulfide Isomerase Accelerates the Folding of Cone Snail Toxins
title Ero1-Mediated Reoxidation of Protein Disulfide Isomerase Accelerates the Folding of Cone Snail Toxins
title_full Ero1-Mediated Reoxidation of Protein Disulfide Isomerase Accelerates the Folding of Cone Snail Toxins
title_fullStr Ero1-Mediated Reoxidation of Protein Disulfide Isomerase Accelerates the Folding of Cone Snail Toxins
title_full_unstemmed Ero1-Mediated Reoxidation of Protein Disulfide Isomerase Accelerates the Folding of Cone Snail Toxins
title_short Ero1-Mediated Reoxidation of Protein Disulfide Isomerase Accelerates the Folding of Cone Snail Toxins
title_sort ero1-mediated reoxidation of protein disulfide isomerase accelerates the folding of cone snail toxins
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6275033/
https://www.ncbi.nlm.nih.gov/pubmed/30384459
http://dx.doi.org/10.3390/ijms19113418
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