Peptide binding to MHC class I proteins measured with a novel fluorescent technique.
An N-dansylated peptide derived from the Plasmodium berghei circumsporozoite protein (PbCS 253-260) bound in an allele-specific manner to a single-chain Kd molecule (SC-Kd), and its binding resulted in significant fluorescent enhancement. The binding kinetics of unlabelled peptides could be determined by pre-incubating dansylated PbCS with a concentrated suspension of SC-Kd, and then diluting this mixture in the presence of unmodified peptide. The time-dependence of the ensuing fluorescence decrease could be fitted to a single-exponential, which gave an association rate constant of 77 M-1s-1 for unlabelled PbCS.
Ojcius
D M
DM
Institut Pasteur, I.N.S.E.R.M. U. n. 277, Unité de Biologie moléculaire du Gène, Paris.
Abastado
J P
JP
Godeau
F
F
Kourilsky
P
P
D000818:Animals; D003619:Dansyl Compounds / Q000378:metabolism*; D005453:Fluorescence; D015395:Histocompatibility Antigens Class I / Q000378:metabolism*; D066298:In Vitro Techniques; D008722:Methods; D010455:Peptides / Q000378:metabolism*; D010962:Plasmodium berghei / Q000737:chemistry; D011485:Protein Binding
D003619:Dansyl Compounds; D015395:Histocompatibility Antigens Class I; D010455:Peptides