A quantitative assay of peptide-dependent class I assembly.
We have developed a quantitative assay for the measurement of class I assembly induced by peptide. We have applied this assay to H-2Db, Kb and HLA-A2.1 with a panel of 49 overlapping peptides derived from HIV-1 gag protein. We find that the effects of peptide on assembly form a continuous distribution. By defining positives as those that increase the concentration of folded heavy chains more than three standard deviations from the control we show that 7/48 bind A2.1, 11/49 bind Db and 7/47 bind Kb. The assembly assay contrasts with solid-phase assays in being more discriminating (fewer peptides binding any given class I molecule), and showing less overlap in the patterns of peptides bound by the three class I molecules.
Elvin
J
J
Institute of Molecular Medicine, Headington, Oxford, Great Britain.
Cerundolo
V
V
Elliott
T
T
Townsend
A
A
D000595:Amino Acid Sequence; D004591:Electrophoresis, Polyacrylamide Gel; D015683:Gene Products, gag / Q000502:physiology*; D006183:H-2 Antigens / Q000096:biosynthesis; D015789:HLA-A2 Antigen / Q000096:biosynthesis; D015395:Histocompatibility Antigens Class I / Q000096:biosynthesis*; D006801:Humans; D007143:Immunoglobulin Heavy Chains / Q000378:metabolism; D008223:Lymphoma / Q000276:immunology; D008969:Molecular Sequence Data; D011233:Precipitin Tests; D014407:Tumor Cells, Cultured; D001613:beta 2-Microglobulin / Q000378:metabolism
D015683:Gene Products, gag; D006183:H-2 Antigens; D015789:HLA-A2 Antigen; D015395:Histocompatibility Antigens Class I; D007143:Immunoglobulin Heavy Chains; D001613:beta 2-Microglobulin