More efficient peptide binding to MHC class II molecules during cathepsin B digestion of Ii than after Ii release.
The binding of a T cell-presented peptide to MHC class II alpha,beta chains occurs as a concurrent process with the release of the associated invariant chain (Ii) by cathepsin B. Ii was digested by cathepsin B from solubilized, MHC class II alpha,beta,Ii complexes in the presence of N-hydroxysuccinimidyl-4-azidobenzoate-conjugated, 125I-labeled, influenza virus matrix (18-29) peptide. The peptide was crosslinked where it became bound. This HLA-DR1-restricted peptide bound about three times more efficiently to class II alpha,beta chains of DR1-positive B cells when present during cathepsin B digestion of Ii than when added afterward, also at pH 5.0. Binding was competed by similarly DR-restricted peptides. Cathepsin D cleaved Ii but did not enhance peptide binding. However, a trace level of cathepsin D, added to the assay for peptide binding in the presence of cathepsin B, further enhanced peptide binding about three times. These experiments support an hypothesis for the staged release of Ii fragments by cathepsin D and cathepsin B, catalyzing at one point the insertion of a peptide into the antigen binding site formed by class II alpha and beta chains.
Daibata
M
M
Department of Pharmacology, University of Massachusetts Medical School, Worcester 01655.
Xu
M
M
Humphreys
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Reyes
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D000595:Amino Acid Sequence; D015703:Antigens, CD / Q000378:metabolism*; D000944:Antigens, Differentiation, B-Lymphocyte / Q000378:metabolism*; D000956:Antigens, Viral / Q000378:metabolism*; D001667:Binding, Competitive; D002401:Cathepsin B / Q000494:pharmacology*; D015798:HLA-DR1 Antigen / Q000378:metabolism; D000949:Histocompatibility Antigens Class II / Q000187:drug effects* / Q000378:metabolism*; D006801:Humans; D008969:Molecular Sequence Data; D009975:Orthomyxoviridae / Q000276:immunology; D010446:Peptide Fragments / Q000378:metabolism; D011485:Protein Binding; D014763:Viral Matrix Proteins / Q000378:metabolism
D015703:Antigens, CD; D000944:Antigens, Differentiation, B-Lymphocyte; D000956:Antigens, Viral; D015798:HLA-DR1 Antigen; D000949:Histocompatibility Antigens Class II; D010446:Peptide Fragments; D014763:Viral Matrix Proteins; C045084:invariant chain; D002401:Cathepsin B