Direct binding of peptide to empty MHC class I molecules on intact cells and in vitro.
MHC class I molecules devoid of peptide are expressed on the cell surface of the mouse mutant lymphoma cell line RMA-S upon culture at reduced temperature. Empty class I molecules are thermolabile at the cell surface and in detergent lysates, but can be stabilized by the addition of presentable peptide; peptide binding appears to be a rapid process. Furthermore, class I molecules on the surface of RMA-S (H-2b haplotype) cells cultured at 26 degrees C can efficiently and specifically bind iodinated peptide presented by H-2Kb. Binding of iodinated peptide is also observed at a lower level for nonmutant cells (RMA) cultured at 26 degrees C. These experiments underscore the role for peptide in maintenance of the structure of class I molecules and, more importantly, provide two assay systems to study the interactions of peptides with MHC class I molecules independent of the availability of T cells that recognize a particular peptide-MHC class I complex.
Schumacher
T N
TN
The Netherlands Cancer Institute, Amsterdam.
Heemels
M T
MT
Neefjes
J J
JJ
Kast
W M
WM
Melief
C J
CJ
Ploegh
H L
HL
D000818:Animals; D002460:Cell Line; D002462:Cell Membrane / Q000276:immunology; D004591:Electrophoresis, Polyacrylamide Gel; D006183:H-2 Antigens / Q000276:immunology; D015395:Histocompatibility Antigens Class I / Q000276:immunology* / Q000302:isolation & purification; D007124:Immunoenzyme Techniques; D051379:Mice; D010455:Peptides / Q000138:chemical synthesis; D011485:Protein Binding
D006183:H-2 Antigens; D015395:Histocompatibility Antigens Class I; D010455:Peptides