A three-domain T cell receptor is biologically active and specifically stains cell surface MHC/peptide complexes.
We have expressed in bacteria a single-chain T cell receptor (scTCR) with specificity for an HIV gp120-derived peptide bound to the murine MHC-I molecule, H-2Dd. This scTCR consists of V alpha covalently linked to the VbetaCbeta domains that was solubilized, refolded, and purified in high yield. Specific binding of the scTCR to MHC/peptide complexes was demonstrated by surface plasmon resonance, with a Kd of 2 to 8 x 10(-6) M. This scTCR specifically inhibited T cell activation, and stained cell surface MHC/peptide complexes as measured by cytofluorimetry. The preservation of binding specificity by such a three-domain scTCR suggests that this structure is sufficient for specific MHC/peptide recognition and that this strategy will be of general use as applied to other TCR.
Plaksin
D
D
Laboratory of Immunology, National Institute of Allergy and Infectious Diseases, Bethesda, MD 20892, USA.
Polakova
K
K
McPhie
P
P
Margulies
D H
DH
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry; D000918:Antibody Specificity; D000937:Antigen-Antibody Reactions; D001483:Base Sequence; D002942:Circular Dichroism; D000939:Epitopes / Q000737:chemistry* / Q000276:immunology; D006183:H-2 Antigens / Q000737:chemistry*; D006825:Hybridomas / Q000276:immunology; D007136:Immunoglobulins / Q000737:chemistry; D008213:Lymphocyte Activation; D051379:Mice; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry* / Q000276:immunology* / Q000502:physiology; D011485:Protein Binding / Q000276:immunology; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D011948:Receptors, Antigen, T-Cell / Q000096:biosynthesis / Q000737:chemistry* / Q000276:immunology; D013194:Staining and Labeling; D013601:T-Lymphocytes / Q000276:immunology
D000911:Antibodies, Monoclonal; D000939:Epitopes; D006183:H-2 Antigens; D007136:Immunoglobulins; D010455:Peptides; D011948:Receptors, Antigen, T-Cell