Characteristics of endogenous peptides eluted from the class I MHC molecule HLA-B7 determined by mass spectrometry and computer modeling.
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.
Huczko
E L
EL
Beirne Carter Center for Immunology Research, University of Virginia, Charlottesville 22901.
Bodnar
W M
WM
Benjamin
D
D
Sakaguchi
K
K
Zhu
N Z
NZ
Shabanowitz
J
J
Henderson
R A
RA
Appella
E
E
Hunt
D F
DF
Engelhard
V H
VH
D000595:Amino Acid Sequence; D003196:Computer Graphics; D003198:Computer Simulation; D017124:Conserved Sequence; D015234:HLA-A Antigens / Q000737:chemistry; D015793:HLA-B7 Antigen / Q000737:chemistry*; D013058:Mass Spectrometry; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry*
D015234:HLA-A Antigens; D015793:HLA-B7 Antigen; D010455:Peptides