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Protein disulfide-isomerase interacts with a substrate protein at all stages along its folding pathway
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Irvine, Alistair, Wallis, A. Katrine, Sanghera, Narinder, Rowe, Michelle L., Ruddock, Lloyd W., Howard, Mark J., Williamson, Richard A., Blindauer, Claudia A. and Freedman, R. B. (2014) Protein disulfide-isomerase interacts with a substrate protein at all stages along its folding pathway. PLoS One, Volume 9 (Number 1). Article number e82511. doi:10.1371/journal.pone.0082511 ISSN 1932-6203.
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Text (http://creativecommons.org/licenses/by-nc-sa/4.0/)
WRAP_2_journal.pone.0082511.pdf - Published Version Download (1519Kb) | Preview |
Official URL: http://dx.doi.org/10.1371/journal.pone.0082511
Abstract
In contrast to molecular chaperones that couple protein folding to ATP hydrolysis, protein disulfide-isomerase (PDI) catalyzes protein folding coupled to formation of disulfide bonds (oxidative folding). However, we do not know how PDI distinguishes folded, partly-folded and unfolded protein substrates. As a model intermediate in an oxidative folding pathway, we prepared a two-disulfide mutant of basic pancreatic trypsin inhibitor (BPTI) and showed by NMR that it is partly-folded and highly dynamic. NMR studies show that it binds to PDI at the same site that binds peptide ligands, with rapid binding and dissociation kinetics; surface plasmon resonance shows its interaction with PDI has a Kd of ca. 10−5 M. For comparison, we characterized the interactions of PDI with native BPTI and fully-unfolded BPTI. Interestingly, PDI does bind native BPTI, but binding is quantitatively weaker than with partly-folded and unfolded BPTI. Hence PDI recognizes and binds substrates via permanently or transiently unfolded regions. This is the first study of PDI's interaction with a partly-folded protein, and the first to analyze this folding catalyst's changing interactions with substrates along an oxidative folding pathway. We have identified key features that make PDI an effective catalyst of oxidative protein folding – differential affinity, rapid ligand exchange and conformational flexibility.
Item Type: | Journal Article | ||||
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Subjects: | Q Science > QD Chemistry | ||||
Divisions: | Faculty of Science, Engineering and Medicine > Science > Chemistry Faculty of Science, Engineering and Medicine > Science > Life Sciences (2010- ) |
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Library of Congress Subject Headings (LCSH): | Protein disulfide isomerase , Aprotinin | ||||
Journal or Publication Title: | PLoS One | ||||
Publisher: | Public Library of Science | ||||
ISSN: | 1932-6203 | ||||
Official Date: | 2014 | ||||
Dates: |
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Volume: | Volume 9 | ||||
Number: | Number 1 | ||||
Page Range: | Article number e82511 | ||||
DOI: | 10.1371/journal.pone.0082511 | ||||
Status: | Peer Reviewed | ||||
Publication Status: | Published | ||||
Access rights to Published version: | Open Access (Creative Commons) | ||||
Date of first compliant deposit: | 26 December 2015 | ||||
Date of first compliant Open Access: | 26 December 2015 | ||||
Funder: | Engineering and Physical Sciences Research Council (EPSRC), Biotechnology and Biological Sciences Research Council (Great Britain) (BBSRC), Wellcome Trust (London, England), Royal Society (Great Britain) | ||||
Grant number: | BB/D017807 (BBSRC) ; 093125/Z/10Z, 047795/Z/96 (WT) |
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