Laboratory evolution of one disulfide isomerase to resemble another

Hiniker, Annie, Ren, Guoping, Heras, Begoña, Zheng, Ying, Laurinec, Stephanie, Jobson, Richard W., Stuckey, Jeanne A., Martin, Jennifer L. and Bardwell, James C. A. (2007) Laboratory evolution of one disulfide isomerase to resemble another. Proceedings of The National Academy of Sciences of The United States of America, 104 28: 11670-11675. doi:10.1073/pnas.0704692104


Author Hiniker, Annie
Ren, Guoping
Heras, Begoña
Zheng, Ying
Laurinec, Stephanie
Jobson, Richard W.
Stuckey, Jeanne A.
Martin, Jennifer L.
Bardwell, James C. A.
Title Laboratory evolution of one disulfide isomerase to resemble another
Journal name Proceedings of The National Academy of Sciences of The United States of America   Check publisher's open access policy
ISSN 0027-8424
1091-6490
Publication date 2007-07-10
Sub-type Article (original research)
DOI 10.1073/pnas.0704692104
Open Access Status Not Open Access
Volume 104
Issue 28
Start page 11670
End page 11675
Total pages 6
Place of publication Washington, D.C., U. S. A.
Publisher National Academy of Sciences
Collection year 2008
Language eng
Subject C1
270199 Biochemistry and Cell Biology not elsewhere classified
780105 Biological sciences
Formatted abstract
It is often difficult to determine which of the sequence and structural differences between divergent members of multigene families are functionally important. Here we use a laboratory evolution approach to determine functionally important structural differences between two distantly related disulfide isomerases, DsbC and DsbG from Escherichia coli. Surprisingly, we found single amino acid substitutions in DsbG that were able to complement dsbC in vivo and have more DsbC-Iike isomerase activity in vitro. Crystal structures of the three strongest point mutants, DsbG K113E, DsbG V216M, and DsbG T200M, reveal changes in highly surface-exposed regions that cause DsbG to more closely resemble the distantly related DsbC. In this case, laboratory evolution appears to have taken a direct route to allow one protein family member to complement another, with single substitutions apparently bypassing much of the need for multiple changes that took place over ~0.5 billion years of evolution. Our findings suggest that, for these two proteins at least, regions important in determining functional differences may represent only a tiny fraction of the overall protein structure.
© 2007 by The National Academy of Sciences of the USA
Keyword Multidisciplinary Sciences
Chaperone
Protein folding
Directed evolution
Escherichia-coli
In-vivo
DsbC
Oxidoreductase
Vitro
Isomerization
Pathway
Oxidase
Enzyme
Domain
DsbG
Q-Index Code C1
Q-Index Status Confirmed Code

 
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Created: Mon, 18 Feb 2008, 14:23:27 EST