The structure of the bacterial oxidoreductase enzyme DsbA in complex with a peptide reveals a basis for substrate specificity in the catalytic cycle of DsbA enzymes

Paxman, Jason J., Borg, Natalie A., Horne, James, Thompson, Philip E., Chin, Yanni, Sharma, Pooja, Simpson, Jamie S., Wielens, Jerome, Piek, Susannah, Kahler, Charlene M., Sakellaris, Harry, Pearce, Mary, Bottomley, Stephen P., Rossjohn, Jamie and Scanlon, Martin J. (2009) The structure of the bacterial oxidoreductase enzyme DsbA in complex with a peptide reveals a basis for substrate specificity in the catalytic cycle of DsbA enzymes. Journal of Biological Chemistry, 284 26: 17835-17845. doi:10.1074/jbc.M109.011502

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Author Paxman, Jason J.
Borg, Natalie A.
Horne, James
Thompson, Philip E.
Chin, Yanni
Sharma, Pooja
Simpson, Jamie S.
Wielens, Jerome
Piek, Susannah
Kahler, Charlene M.
Sakellaris, Harry
Pearce, Mary
Bottomley, Stephen P.
Rossjohn, Jamie
Scanlon, Martin J.
Title The structure of the bacterial oxidoreductase enzyme DsbA in complex with a peptide reveals a basis for substrate specificity in the catalytic cycle of DsbA enzymes
Journal name Journal of Biological Chemistry   Check publisher's open access policy
ISSN 0021-9258
1083-351X
Publication date 2009-06
Sub-type Article (original research)
DOI 10.1074/jbc.M109.011502
Open Access Status File (Publisher version)
Volume 284
Issue 26
Start page 17835
End page 17845
Total pages 11
Place of publication American Society for Biochemistry and Molecular Biology
Publisher Bethesda, MD, United States
Language eng
Abstract Oxidative protein folding in Gram-negative bacteria results in the formation of disulfide bonds between pairs of cysteine residues. This is a multistep process in which the dithiol-disulfide oxidoreductase enzyme, DsbA, plays a central role. The structure of DsbA comprises an all helical domain of unknown function and a thioredoxin domain, where active site cysteines shuttle between an oxidized, substrate-bound, reduced form and a DsbB-bound form, where DsbB is a membrane protein that reoxidizes DsbA. Most DsbA enzymes interact with a wide variety of reduced substrates and show little specificity. However, a number of DsbA enzymes have now been identified that have narrow substrate repertoires and appear to interact specifically with a smaller number of substrates. The transient nature of the DsbA-substrate complex has hampered our understanding of the factors that govern the interaction of DsbA enzymes with their substrates. Here we report the crystal structure of a complex between Escherichia coli DsbA and a peptide with a sequence derived from a substrate. The binding site identified in the DsbA-peptide complex was distinct from that observed for DsbB in the DsbA-DsbB complex. The structure revealed details of the DsbA-peptide interaction and suggested a mechanism by which DsbA can simultaneously show broad specificity for substrates yet exhibit specificity for DsbB. This mode of binding was supported by solution nuclear magnetic resonance data as well as functional data, which demonstrated that the substrate specificity of DsbA could be modified via changes at the binding interface identified in the structure of the complex
Keyword Disulfide Bond Formation
Staphylococcus-Aureus Dsba
III Secretion System
Escherichia-Coli
Neisseria-Meningitidis
Crystal-Structure
Vibrio-Cholerae
Pseudomonas-Aeruginosa
Shigella-Flexneri
Virulence Factors
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status Non-UQ

Document type: Journal Article
Sub-type: Article (original research)
Collection: Institute for Molecular Bioscience - Publications
 
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Created: Thu, 16 Aug 2012, 10:18:01 EST by Susan Allen on behalf of Institute for Molecular Bioscience