Interaction of antigenic peptides with MHC class I molecules on living cells studied by photoaffinity labeling.

Details

Serval ID
serval:BIB_5AC5C286F11E
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Interaction of antigenic peptides with MHC class I molecules on living cells studied by photoaffinity labeling.
Journal
Journal of immunology
Author(s)
Luescher I.F., Lóez J.A., Malissen B., Cerottini J.C.
ISSN
0022-1767
Publication state
Published
Issued date
1992
Peer-reviewed
Oui
Volume
148
Number
4
Pages
1003-1011
Language
english
Notes
Publication types: Journal Article
Publication Status: ppublish
Abstract
Using a direct binding assay based on photoaffinity labeling, we have studied the interaction of antigenic peptides with murine MHC class I molecules on living cells. Photoreactive derivatives were prepared by N-terminal amidation with iodo, 4-azido salicylic acid of the Kd restricted Plasmodium berghei circumsporozoite (P.b. CS) peptide 253-260 (YIPSAEKI) and the Db-restricted Adenovirus 5 early region 1A (Ad5 E1A) peptide 234-243 (SGPSNTPPEI). As assessed in functional competition experiments, both peptide derivatives retained the specific binding activity of the parental peptides for Kd or Dd, respectively. The P.b. CS photoprobe specifically labeled Kd molecules on P815 (H-2d) cells, but failed to label RMA (H-2b) cells. Conversely, the Ad5 E1A photoprobe specifically labeled Db molecules on RMA cells, but failed to label P815 cells. When the two photoprobes were tested on a panel of Con A-activated spleen cells expressing 10 different H-2 haplotypes, significant photoaffinity labeling was observed only on H-2d cells with the P.b. CS photoprobe and on H-2b cells with the Ad5 E1A photoprobe. Labeling of cell-associated Kd or Db molecules with the photoprobes was specifically inhibited by antigenic peptides known to be presented by the same class I molecule. Photoaffinity labeling of Kd with the P.b. CS photoprobe was used to study the dynamics of peptide binding on living P815 cells. Binding increased steadily with the incubation period (up to 8 h) at 37 degrees C and at ambient temperature, but was greatly reduced (greater than 95%) at 0 to 4 degrees C or in the presence of ATP synthesis inhibitors. The magnitude of the labeling was twofold higher at room temperature than at 37 degrees C. In contrast, binding to isolated Kd molecules in solution rapidly reached maximal binding, particularly at 37 degrees C. Dissociation of the photoprobe from either cell-associated or soluble Kd molecules was similar, with a half time of approximately 1 h at 37 degrees C, whereas the complexes were long-lived at 4 degrees C in both instances.
Keywords
Adenosine Triphosphate/physiology, Affinity Labels/metabolism, Alleles, Amino Acid Sequence, Animals, H-2 Antigens/metabolism, Histocompatibility Antigens Class I/analysis, Histocompatibility Antigens Class I/metabolism, Mice, Mice, Inbred C57BL, Molecular Sequence Data, Peptide Fragments/metabolism, T-Lymphocytes, Cytotoxic/immunology, Temperature
Pubmed
Web of science
Create date
28/01/2008 11:20
Last modification date
20/08/2019 14:13
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