Peptide specificity of RT1-A1(c), an inhibitory rat major histocompatibility complex class I natural killer cell ligand.
The rat major histocompatibility complex class Ia allelomorph RT1-A1(c) is a potent ligand for the recently identified inhibitory rLy-49 receptor, STOK-2. With the ultimate objective of studying the interactions of these molecules using structural and functional methods, we undertook a detailed study of its peptide specificity. The study revealed that designing an "ideal peptide" by choosing the most abundant residues in the "binding motif" obtained by pool sequencing does not necessarily yield an optimal binding peptide. For RT1-A1(c), as many as four positions, P2, P4, P5, and P9, were detected as putative anchors. Since this molecule displays a preference for highly hydrophobic peptides, we tested binding of peptides derived from the known leader peptide sequences of other rat histocompatibility complex class I molecules. One such peptide, found to bind well, requiring 1.6 microm peptide to achieve 50% stabilization, was searched for in vivo. Natural RT1-A1(c) binding peptides were purified from rat splenocytes and characterized by mass spectrometry using a combined matrix-assisted laser desorption ionization/time-of-flight and quadrupole time-of-flight approach. Results showed that the signal sequence-derived peptide was not detectable in the purified peptide pool, which was composed of a complex spectrum of peptides. Seven of these self-peptides were successfully sequenced.
Stevens
J
J
Molecular Immunology Programme, Babraham Institute, Cambridge CB2 4AT, UK. jstevens@scripps.edu
Jones
R C
RC
Bordoli
R S
RS
Trowsdale
J
J
Gaskell
S J
SJ
Butcher
G W
GW
Joly
E
E
D000818:Animals; D000950:Antigens, Ly; D001665:Binding Sites; D006649:Histocompatibility Antigens / Q000276:immunology* / Q000378:metabolism; D007694:Killer Cells, Natural / Q000276:immunology* / Q000378:metabolism; D037181:Lectins, C-Type; D008024:Ligands; D008562:Membrane Glycoproteins / Q000276:immunology* / Q000378:metabolism; D010455:Peptides / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D051381:Rats; D011971:Receptors, Immunologic / Q000276:immunology / Q000378:metabolism; D055630:Receptors, NK Cell Lectin-Like
D000950:Antigens, Ly; D006649:Histocompatibility Antigens; D037181:Lectins, C-Type; D008024:Ligands; D008562:Membrane Glycoproteins; D010455:Peptides; D011971:Receptors, Immunologic; D055630:Receptors, NK Cell Lectin-Like; C038353:histocompatibility antigens RT, rat