A synthetic IgG-binding domain based on staphylococcal protein A

Protein Eng. 1987 Feb-Mar;1(2):107-13. doi: 10.1093/protein/1.2.107.

Abstract

A synthetic IgG-binding domain based on staphylococcal protein A was designed with the aid of sequence comparisons and computer graphic analysis. A strategy, utilizing non-palindromic restriction sites, was used to overcome the difficulties of introducing site-specific changes into the repetitive gene. A single mutagenized gene fragment was polymerized to different multiplicities, and the different gene products were expressed in Escherichia coli. Using this scheme, protein A-like proteins composed of different numbers of IgG-binding domains were produced. These domains were changed to lack asparagine--glycine dipeptide sequences as well as methionine residues and are thus, in contrast to native protein A, resistant to treatment with hydroxylamine and cyanogen bromide.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Base Sequence
  • Binding Sites
  • Cloning, Molecular
  • DNA / genetics
  • Immunoglobulin G* / metabolism
  • Models, Molecular
  • Molecular Sequence Data
  • Protein Conformation
  • Protein Engineering
  • Repetitive Sequences, Nucleic Acid
  • Staphylococcal Protein A* / genetics
  • Staphylococcal Protein A* / metabolism

Substances

  • Immunoglobulin G
  • Staphylococcal Protein A
  • DNA