TY - JOUR
T1 - Rapid acidification and alkylation
T2 - redox analysis of the MHC class I pathway
AU - Powis, Simon J
AU - Nesbeth, Darren
AU - Lenart, Izabela
AU - Fussell, Helen
AU - Lamb, Tracey
AU - Gould, Keith
AU - Antoniou, Antony N
PY - 2009/1/1
Y1 - 2009/1/1
N2 - 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.
AB - 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.
KW - Alkylation
KW - Cysteine/chemistry
KW - HLA-A2 Antigen/chemistry
KW - HLA-B27 Antigen/chemistry
KW - Humans
KW - Hydrogen-Ion Concentration
KW - Oxidation-Reduction
KW - Oxidoreductases/chemistry
KW - Stilbenes/chemistry
KW - Sulfonic Acids/chemistry
KW - Trichloroacetic Acid/chemistry
U2 - 10.1016/j.jim.2008.09.006
DO - 10.1016/j.jim.2008.09.006
M3 - Article
C2 - 18838077
SN - 0022-1759
VL - 340
SP - 81
EP - 85
JO - Journal of Immunological Methods
JF - Journal of Immunological Methods
IS - 1
ER -