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Protein Eng. 1987 Feb-Mar;1(2):107-13.

A synthetic IgG-binding domain based on staphylococcal protein A.

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Department of Biochemistry and Biotechnology, Royal Institute of Technology, Stockholm, Sweden.


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.

[Indexed for MEDLINE]

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