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The Journal of Immunology, Vol 151, Issue 5 2572-2587, Copyright © 1993 by American Association of Immunologists

ARTICLES

Characteristics of endogenous peptides eluted from the class I MHC molecule HLA-B7 determined by mass spectrometry and computer modeling

EL Huczko, WM Bodnar, D Benjamin, K Sakaguchi, NZ Zhu, J Shabanowitz, RA Henderson, E Appella, DF Hunt and VH Engelhard
Beirne Carter Center for Immunology Research, University of Virginia, Charlottesville 22901.

Microcapillary HPLC electrospray ionization tandem mass spectrometry was used to sequence 15 peptides eluted from HLA-B7. Sequence alignment implicated four peptide positions in specific interactions with the class I molecule, and their importance was confirmed using synthetic peptides. Because no crystal structure for HLA-B7 was available, computer-assisted modeling was used to understand novel aspects of peptide binding specificity and to accurately predict the effect of defined changes in peptide structure. The results demonstrate that mass- spectrometric sequencing coupled with computer-assisted modeling can be used in the absence of a crystal structure to make accurate predictions concerning requirements for peptide binding to class I molecules. These techniques may be valuable to predict or engineer T cell epitopes.


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