Testing the importance of each residue in a HLA-B27-binding peptide using monoclonal antibodies.
When a peptide derived from histone 3.3 was incubated with mouse L cells transfected with HLA-B27, the cells became highly reactive with Ye-2, an anti-HLA-B27 mAb. The critical residues were analyzed by testing analogues in which each of the nine residues in the peptide was consecutively substituted by 19 other amino acids. The conclusions were separately verified using a different HLA-B27-positive cell line. The ability of some of these peptides to bind to HLA-B27 was also assayed by their ability to stabilize HLA-B27 in a mutant cell line which required HLA-B27-binding peptides to express HLA-B27 at 37 degrees C. These experiments showed that in P4, P5, P6, P7, P8, and P9, all 20 different amino acids could be substituted without eliminating the ability of the analogues to bind to HLA-B27. The residues which were responsible for the HLA-B27-peptide complex reacting with the Ye-2 antibody were P8 and P9. The latter might mediate its effect by altering either the surface conformation of the closely associated HLA-B27 heavy chain or the conformation of the peptide itself.
Fukazawa
T
T
Department of Medicine, University of California, Los Angeles 90024.
Wang
J
J
Huang
F
F
Wen
J
J
Tyan
D
D
Williams
K M
KM
Raybourne
R B
RB
Yu
D T
DT
D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000276:immunology*; D015796:HLA-B27 Antigen / Q000378:metabolism*; D006360:Heat-Shock Proteins / Q000737:chemistry / Q000276:immunology; D006657:Histones / Q000737:chemistry / Q000276:immunology; D006801:Humans; D066298:In Vitro Techniques; D008969:Molecular Sequence Data; D010455:Peptides / Q000276:immunology* / Q000378:metabolism
D000911:Antibodies, Monoclonal; D015796:HLA-B27 Antigen; D006360:Heat-Shock Proteins; D006657:Histones; D010455:Peptides