Binding of radioiodinated influenza virus peptides to class I MHC molecules and to other cellular proteins as analyzed by gel filtration and photoaffinity labeling
Graduate School of Biomedical Sciences; Department of Pharmacology
Medical Subject Headings
Affinity Labels; Amino Acid Sequence; Antigens, Viral; Binding, Competitive; Cell Line; Chromatography, Gel; HLA-B Antigens; Histocompatibility Antigens Class I; Humans; Membrane Proteins; Microsomes; Molecular Sequence Data; Nucleoproteins; Orthomyxoviridae; Peptides; Viral Matrix Proteins
Life Sciences | Medicine and Health Sciences
In order to determine how T cell-presented peptides associate with the antigen binding sites (desetopes) of class I major histocompatibility complex (MHC) molecules and how they might be scavenged from an endogenous processing pathway for transfer to those molecules, we characterized the binding of two synthetic peptides restricted by HLA-B37 or HLA-A2 to class I MHC molecules and to cellular proteins of histotyped cell lines, by gel filtration and photo-affinity labeling techniques. In gel filtration binding studies, each peptide associated with immunopurified class I MHC molecules from cells with its restricting, histotype, but little was bound to class I MHC molecules from cells without the restricting histotype and none was bound to bovine serum albumin. After crosslinkage of a radioiodinated photoreactive derivative of influenza virus nucleoprotein peptide NP(336-355Y) and immunoprecipitations with antibodies to class I MHC molecules, that peptide was found to bind to immunopurified class I MHC molecules from HLA-B37+ but not HLA-B37- cells. Binding of the [125I]NP peptide increased from 6 to 12 hr of incubation and was competed by unlabeled, NP peptide but not by HLA-A2-restricted, influenza virus matrix MA(57-73). The principal microsomal membrane proteins binding [125I]NP were about 65, 45 and 33 kD.
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Citation: Mol Immunol. 1991 Apr-May;28(4-5):341-8.
Reyes, Victor E.; Lu, Shan; and Humphreys, Robert E., "Binding of radioiodinated influenza virus peptides to class I MHC molecules and to other cellular proteins as analyzed by gel filtration and photoaffinity labeling" (1991). GSBS Student Publications. 1041.