Rapid acidification and alkylation: redox analysis of the MHC class I pathway.
The technique of rapid acidification and alkylation can be used to characterise the redox status of oxidoreductases, and to determine numbers of free cysteine residues within substrate proteins. We have previously used this method to analyse interacting components of the MHC class I pathway, namely ERp57 and tapasin. Here, we have applied rapid acidification/alkylation as a novel approach to analysing the redox status of MHC class I molecules. This analysis of the redox status of the MHC class I molecules HLA-A2 and HLA-B27, which is strongly associated with a group of inflammatory arthritic disorders referred to as Spondyloarthropathies, revealed structural and conformational information. We propose that this assay provides a useful tool in the study of in vivo MHC class I structure.
Powis
Simon J
SJ
Bute Medical School, University of St. Andrews, Fife, Scotland, KY16 9TS, United Kingdom.
Nesbeth
Darren
D
Lenart
Izabela
I
Fussell
Helen
H
Lamb
Tracey
T
Gould
Keith
K
Antoniou
Antony N
AN
D000478:Alkylation; D003545:Cysteine / Q000737:chemistry / Q000276:immunology / Q000378:metabolism; D015789:HLA-A2 Antigen / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D015796:HLA-B27 Antigen / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D010084:Oxidation-Reduction; D010088:Oxidoreductases / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D013267:Stilbenes / Q000737:chemistry; D013451:Sulfonic Acids / Q000737:chemistry; D014238:Trichloroacetic Acid / Q000737:chemistry
D015789:HLA-A2 Antigen; D015796:HLA-B27 Antigen; D013267:Stilbenes; D013451:Sulfonic Acids; C079532:4-acetamido-4'-maleimidylstilbene-2,2'-disulfonic acid; D014238:Trichloroacetic Acid; D010088:Oxidoreductases; D003545:Cysteine