Rapid acidification and alkylation: redox analysis of the MHC class I pathway

Simon J Powis, Darren Nesbeth, Izabela Lenart, Helen Fussell, Tracey Lamb, Keith Gould, Antony N Antoniou

Research output: Contribution to journalArticlepeer-review

3 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)81-5
Number of pages5
JournalJournal of Immunological Methods
Volume340
Issue number1
DOIs
Publication statusPublished - 1 Jan 2009

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