Open AccessBinding of soluble natural ligands to a soluble human T-cell receptor fragment produced in Escherichia coli.
Author(s) -
Katherine L. Hilyard,
Hugh Reyburn,
Shan Chung,
John I. Bell,
Jack L. Strominger
Publication year - 1994
Publication title -
proceedings of the national academy of sciences of the united states of america
Language(s) - English
Resource type - Journals
eISSN - 1091-6490
pISSN - 0027-8424
DOI - 10.1073/pnas.91.19.9057
Subject(s) - escherichia coli , enterotoxin , recombinant dna , biochemistry , chemistry , peptide , inclusion bodies , biology , fusion protein , microbiology and biotechnology , receptor , gene
An Escherichia coli expression system has been developed to produce milligram quantities of the variable domains of a human T-cell receptor from a cytotoxic T cell that recognizes the HLA-A2-influenza matrix peptide complex as a single polypeptide chain. The recombinant protein was purified by metal-chelate chromatography and then refolded in a redox buffer system. The refolded protein was shown to directly bind both Staphylococcus aureus enterotoxin B and the major histocompatibility complex protein-peptide complex using a BIAcore biosensor. Thus this preparation of a single-chain, variable-domain, T-cell receptor fragment can bind both of its natural ligands and some of it is therefore a functional fragment of the receptor molecule.