Energy transfer between two peptides bound to one MHC class II molecule.
The 17-amino acid peptide from chicken ovalbumin, Ova(323-339), was labeled at the amino terminus with fluorescein [FOva(323-339)] and near the carboxyl terminus with Texas Red [AcOva(323-338)KTR]. Fluorescence spectroscopy was carried out on resolved electrophoretic bands on nonreducing polyacrylamide gels derived from incubation mixtures containing major histocompatibility complex (MHC) class II molecules IAd and the FOva(323-339)- and AcOva(323-338)KTR-labeled peptides. Energy transfer between fluorescein and Texas Red was observed in the "floppy" alpha beta heterodimer band, but not in the "compact" alpha beta heterodimer band. Energy transfer was detected between the truncated peptides FOva(323-328)CONH2 and AcOva(331-338)KTR in both the compact alpha beta and floppy alpha beta gel bands. The energy-transfer data suggest that the two binding sites of floppy alpha beta arise from splitting apart a putative large, single binding site region in compact alpha beta.
Tampé
R
R
Stauffer Laboratory for Physical Chemistry, Stanford University, CA 94305.
Clark
B R
BR
McConnell
H M
HM
D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D004735:Energy Transfer; D000949:Histocompatibility Antigens Class II / Q000737:chemistry / Q000378:metabolism*; D066298:In Vitro Techniques; D051379:Mice; D008969:Molecular Sequence Data; D010047:Ovalbumin / Q000737:chemistry; D010455:Peptides / Q000737:chemistry / Q000378:metabolism*; D013050:Spectrometry, Fluorescence; D013329:Structure-Activity Relationship
D000949:Histocompatibility Antigens Class II; D010455:Peptides; D010047:Ovalbumin