PMID- 12220628
OWN - NLM
STAT- MEDLINE
DCOM- 20021108
LR  - 20190621
IS  - 0014-5793 (Print)
IS  - 0014-5793 (Linking)
VI  - 527
IP  - 1-3
DP  - 2002 Sep 11
TI  - Identification of a dominant self-ligand bound to three HLA B44 alleles and the 
      preliminary crystallographic analysis of recombinant forms of each complex.
PG  - 27-32
AB  - A naturally processed and presented ligand that is shared by human leukocyte 
      antigen (HLA) B*4402, B*4403 and B*4405 molecules has been identified in peptides 
      isolated from immunoaffinity purified HLA B44 complexes. This peptide derived 
      from HLA DPalpha residues 46-54, an endogenous product of HLA DP expressed in the 
      cell line Hmy2.C1R, is a prominent peptide in the mass spectra of species 
      isolated as bound peptides from each allele when the three HLA B44 subtypes were 
      introduced as transfected gene products. Recombinant truncated forms of HLA 
      B*4405(1-276), HLA B*4403(1-276), HLA B*4402(1-276) and beta(2)-microglobulin 
      have been prepared as inclusion bodies in Escherichia coli and refolded in the 
      presence of the DPalpha(46-54) peptide and purified by a combination of size 
      exclusion and anion exchange chromatography. This material was determined to be 
      correctly folded based on detection of a conformational epitope recognized by the 
      W6/32 monoclonal antibody. Large, plate-like crystals of the three complexes were 
      produced using polyethylene glycol as the precipitant. All the crystals belong to 
      the space group P2(1)2(1)2(1) with unit cell dimensions of approximately a=51, 
      b=82, c=110 A. The crystals of three B44/DPalpha complexes diffracted to a 
      resolution of 1.9 A or better. For the first time, using this natural, high 
      abundance ligand of the HLA B44 molecules we have successfully expressed and 
      refolded the three HLA B44 molecules and produced crystals amenable to structural 
      studies.
FAU - Macdonald, Whitney
AU  - Macdonald W
AD  - Department of Microbiology and Immunology, University of Melbourne, 3010, 
      Melbourne, Vic., Australia.
FAU - Williams, David S
AU  - Williams DS
FAU - Clements, Craig S
AU  - Clements CS
FAU - Gorman, Jeffery J
AU  - Gorman JJ
FAU - Kjer-Nielsen, Lars
AU  - Kjer-Nielsen L
FAU - Brooks, Andrew G
AU  - Brooks AG
FAU - McCluskey, James
AU  - McCluskey J
FAU - Rossjohn, Jamie
AU  - Rossjohn J
FAU - Purcell, Anthony W
AU  - Purcell AW
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - England
TA  - FEBS Lett
JT  - FEBS letters
JID - 0155157
RN  - 0 (Epitopes)
RN  - 0 (HLA-B Antigens)
RN  - 0 (HLA-B44 Antigen)
RN  - 0 (HLA-DP Antigens)
RN  - 0 (Ligands)
RN  - 0 (Peptides)
RN  - 0 (Recombinant Proteins)
RN  - 0 (beta 2-Microglobulin)
SB  - IM
MH  - Alleles
MH  - Crystallography, X-Ray
MH  - Epitopes
MH  - Escherichia coli/genetics
MH  - HLA-B Antigens/*chemistry/genetics/isolation & purification/*metabolism
MH  - HLA-B44 Antigen
MH  - HLA-DP Antigens/immunology/*metabolism
MH  - Humans
MH  - Ligands
MH  - Peptides/genetics/isolation & purification/metabolism
MH  - Protein Folding
MH  - Recombinant Proteins/chemistry/genetics/immunology
MH  - Tumor Cells, Cultured
MH  - beta 2-Microglobulin/chemistry/genetics/metabolism
EDAT- 2002/09/11 10:00
MHDA- 2002/11/26 04:00
CRDT- 2002/09/11 10:00
PHST- 2002/09/11 10:00 [pubmed]
PHST- 2002/11/26 04:00 [medline]
PHST- 2002/09/11 10:00 [entrez]
AID - S0014579302031496 [pii]
AID - 10.1016/s0014-5793(02)03149-6 [doi]
PST - ppublish
SO  - FEBS Lett. 2002 Sep 11;527(1-3):27-32. doi: 10.1016/s0014-5793(02)03149-6.