title: Binding of soluble natural ligands to a soluble human T-cell receptor fragment produced in Escherichia coli.
abstract: 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.
authors.affiliation: Department of Biochemistry and Molecular Biology, Harvard University, Cambridge, MA 02138.
authors.forename: K L
authors.initials: KL
authors.lastname: Hilyard
authors.forename: H
authors.initials: H
authors.lastname: Reyburn
authors.forename: S
authors.initials: S
authors.lastname: Chung
authors.forename: J I
authors.initials: JI
authors.lastname: Bell
authors.forename: J L
authors.initials: JL
authors.lastname: Strominger
mesh_terms: D000595:Amino Acid Sequence; D015374:Biosensing Techniques; D004768:Enterotoxins / Q000378:metabolism; D004926:Escherichia coli; D015789:HLA-A2 Antigen / Q000378:metabolism*; D006801:Humans; D008024:Ligands; D008969:Molecular Sequence Data; D011485:Protein Binding; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000737:chemistry / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism; D012995:Solubility
chemical_list: D004768:Enterotoxins; D015789:HLA-A2 Antigen; D008024:Ligands; D016693:Receptors, Antigen, T-Cell, alpha-beta; D011994:Recombinant Proteins; C031818:enterotoxin B, staphylococcal