The murine liver-specific nonclassical MHC class I molecule Q10 binds a classical peptide repertoire.
The biological properties of the nonclassical class I MHC molecules secreted into blood and tissue fluids are not currently understood. To address this issue, we studied the murine Q10 molecule, one of the most abundant, soluble class Ib molecules. Mass spectrometry analyses of hybrid Q10 polypeptides revealed that alpha1alpha2 domains of Q10 associate with 8-9 long peptides similar to the classical class I MHC ligands. Several of the sequenced peptides matched intracellularly synthesized murine proteins. This finding and the observation that the Q10 hybrid assembly is TAP2-dependent supports the notion that Q10 groove is loaded by the classical class I Ag presentation pathway. Peptides eluted from Q10 displayed a binding motif typical of H-2K, D, and L ligands. They carried conserved residues at P2 (Gly), P6 (Leu), and Pomega (Phe/Leu). The role of these residues as anchors/auxiliary anchors was confirmed by Ala substitution experiments. The Q10 peptide repertoire was heterogeneous, with 75% of the groove occupied by a multitude of diverse peptides; however, 25% of the molecules bound a single peptide identical to a region of a TCR V beta-chain. Since this peptide did not display enhanced binding affinity for Q10 nor does its origin and sequence suggest that it is functionally significant, we propose that the nonclassical class I groove of Q10 resembles H-2K, D, and L grooves more than the highly specialized clefts of nonclassical class I Ags such as Qa-1, HLA-E, and M3.
Zappacosta
F
F
Laboratory of Allergic Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Rockville, MD 20852, USA.
Tabaczewski
P
P
Parker
K C
KC
Coligan
J E
JE
Stroynowski
I
I
D018528:ATP-Binding Cassette Transporters / Q000502:physiology; D000818:Animals; D002460:Cell Line; D002462:Cell Membrane / Q000276:immunology / Q000378:metabolism; D015395:Histocompatibility Antigens Class I / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D008099:Liver / Q000276:immunology*; D046911:Macromolecular Substances; D051379:Mice; D008809:Mice, Inbred C3H; D008810:Mice, Inbred C57BL; D016688:Mice, Inbred NOD; D009842:Oligopeptides / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism*; D011485:Protein Binding / Q000276:immunology; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D012995:Solubility
D018528:ATP-Binding Cassette Transporters; D015395:Histocompatibility Antigens Class I; D046911:Macromolecular Substances; D009842:Oligopeptides; C032907:Q surface antigens; D011993:Recombinant Fusion Proteins