PMID- 1544922
OWN - NLM
STAT- MEDLINE
DCOM- 19920414
LR  - 20210210
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 267
IP  - 8
DP  - 1992 Mar 15
TI  - Peptide binding to HLA-A2 and HLA-B27 isolated from Escherichia coli. 
      Reconstitution of HLA-A2 and HLA-B27 heavy chain/beta 2-microglobulin complexes 
      requires specific peptides.
PG  - 5451-9
AB  - The specificity of peptide binding by human leukocyte antigen (HLA) class I 
      molecules was investigated in a cell-free direct-binding assay. Peptides were 
      assessed for binding to HLA-A2 and HLA-B27 by measuring the formation of 
      heterotrimeric HLA complexes that consisted of iodinated beta 2-microglobulin, 
      HLA heavy chain fragments isolated from the Escherichia coli cytoplasm, and 
      peptide. In this system, no detectable HLA heavy chain-beta 2-microglobulin 
      complexes were formed unless appropriate peptides were intentionally added to the 
      reconstitution solution. Analysis with monoclonal antibodies demonstrated that 
      these heterotrimeric complexes were correctly folded. Five nonhomologous 
      peptides, known to form complexes with HLA-A2 or HLA-B27 from T-cell functional 
      studies, were tested for their capacity to bind to HLA-A2 and HLA-B27 using the 
      reconstitution assay. Four of the peptides bound to the appropriate class I 
      molecule only. One peptide and some (but not all) substitution analogs of it 
      bound to both HLA-A2 and HLA-B27. The effect of peptide length on binding to 
      HLA-B27 was studied, and it was found that the optimal length was 9 or 10 amino 
      acid residues; however, one peptide that bound to HLA-B27 was 15 amino acids 
      long. All peptides that bound to HLA-B27 in the direct-binding assay also 
      competed with antigenic peptides for binding to HLA-B27 on the surface of intact 
      cells, as determined by a standard cytotoxic T-lymphocyte functional assay. Thus, 
      we conclude that HLA-A2 and HLA-B27 bind distinct but partially overlapping sets 
      of peptides and that, at least in vitro, the assembly of HLA heavy chain-beta 
      2-microglobulin complexes requires specific peptides.
FAU - Parker, K C
AU  - Parker KC
AD  - Biological Resources Branch, National Institute of Allergy and Infectious 
      Diseases, Bethesda, Maryland 20892.
FAU - Carreno, B M
AU  - Carreno BM
FAU - Sestak, L
AU  - Sestak L
FAU - Utz, U
AU  - Utz U
FAU - Biddison, W E
AU  - Biddison WE
FAU - Coligan, J E
AU  - Coligan JE
LA  - eng
PT  - Journal Article
PL  - United States
TA  - J Biol Chem
JT  - The Journal of biological chemistry
JID - 2985121R
RN  - 0 (Antibodies, Monoclonal)
RN  - 0 (HLA-A2 Antigen)
RN  - 0 (HLA-B27 Antigen)
RN  - 0 (Macromolecular Substances)
RN  - 0 (Oligodeoxyribonucleotides)
RN  - 0 (Peptides)
RN  - 0 (Recombinant Proteins)
RN  - 0 (beta 2-Microglobulin)
SB  - IM
MH  - Amino Acid Sequence
MH  - Antibodies, Monoclonal
MH  - Base Sequence
MH  - Cloning, Molecular
MH  - Escherichia coli/*genetics
MH  - HLA-A2 Antigen/genetics/*metabolism
MH  - HLA-B27 Antigen/genetics/*metabolism
MH  - Humans
MH  - Macromolecular Substances
MH  - Molecular Sequence Data
MH  - Oligodeoxyribonucleotides
MH  - Peptides/chemical synthesis/immunology
MH  - Plasmids
MH  - Polymerase Chain Reaction
MH  - Protein Binding
MH  - Recombinant Proteins/metabolism
MH  - beta 2-Microglobulin/*metabolism
EDAT- 1992/03/15 00:00
MHDA- 1992/03/15 00:01
CRDT- 1992/03/15 00:00
PHST- 1992/03/15 00:00 [pubmed]
PHST- 1992/03/15 00:01 [medline]
PHST- 1992/03/15 00:00 [entrez]
AID - S0021-9258(18)42787-1 [pii]
PST - ppublish
SO  - J Biol Chem. 1992 Mar 15;267(8):5451-9.