Skip to main content
placeholder image

Laboratory evolution of one disulfide isomerase to resemble another

Journal Article


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-like 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.

UOW Authors


  •   Hiniker, Annie (external author)
  •   Ren, Guoping (external author)
  •   Heras, Begona (external author)
  •   Zheng, Ying (external author)
  •   Laurinec, Stephanie (external author)
  •   Jobson, Richard (external author)
  •   Stuckey, Jeanne (external author)
  •   Martin, Jennifer (external author)
  •   Bardwell, James (external author)

Publication Date


  • 2007

Citation


  • Hiniker, A., Ren, G., Heras, B., Zheng, Y., Laurinec, S., Jobson, R. W., Stuckey, J. A., Martin, J. L. & Bardwell, J. 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.

Scopus Eid


  • 2-s2.0-34547424808

Number Of Pages


  • 5

Start Page


  • 11670

End Page


  • 11675

Volume


  • 104

Issue


  • 28

Place Of Publication


  • United States

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-like 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.

UOW Authors


  •   Hiniker, Annie (external author)
  •   Ren, Guoping (external author)
  •   Heras, Begona (external author)
  •   Zheng, Ying (external author)
  •   Laurinec, Stephanie (external author)
  •   Jobson, Richard (external author)
  •   Stuckey, Jeanne (external author)
  •   Martin, Jennifer (external author)
  •   Bardwell, James (external author)

Publication Date


  • 2007

Citation


  • Hiniker, A., Ren, G., Heras, B., Zheng, Y., Laurinec, S., Jobson, R. W., Stuckey, J. A., Martin, J. L. & Bardwell, J. 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.

Scopus Eid


  • 2-s2.0-34547424808

Number Of Pages


  • 5

Start Page


  • 11670

End Page


  • 11675

Volume


  • 104

Issue


  • 28

Place Of Publication


  • United States