Conversion of monomeric protein L to an obligate dimer by computational protein design

Proc Natl Acad Sci U S A. 2001 Sep 11;98(19):10687-91. doi: 10.1073/pnas.181354398. Epub 2001 Aug 28.

Abstract

Protein L consists of a single alpha-helix packed on a four-stranded beta-sheet formed by two symmetrically opposed beta-hairpins. We use a computer-based protein design procedure to stabilize a domain-swapped dimer of protein L in which the second beta-turn straightens and the C-terminal strand inserts into the beta-sheet of the partner. The designed obligate dimer contains three mutations (A52V, N53P, and G55A) and has a dissociation constant of approximately 700 pM, which is comparable to the dissociation constant of many naturally occurring protein dimers. The structure of the dimer has been determined by x-ray crystallography and is close to the in silico model.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Bacterial Proteins*
  • DNA-Binding Proteins / chemistry*
  • DNA-Binding Proteins / genetics
  • Dimerization
  • Guanidine
  • Models, Molecular
  • Mutagenesis
  • Protein Denaturation
  • Protein Structure, Secondary

Substances

  • Bacterial Proteins
  • DNA-Binding Proteins
  • L-protein, Peptococcus magnus
  • Guanidine

Associated data

  • PDB/1JML