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

doi:10.1016/j.jim.2008.09.006

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 journal › Article › peer-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 language	English
Pages (from-to)	81-5
Number of pages	5
Journal	Journal of Immunological Methods
Volume	340
Issue number	1
DOIs	https://doi.org/10.1016/j.jim.2008.09.006
Publication status	Published - 1 Jan 2009

Keywords

Alkylation
Cysteine/chemistry
HLA-A2 Antigen/chemistry
HLA-B27 Antigen/chemistry
Humans
Hydrogen-Ion Concentration
Oxidation-Reduction
Oxidoreductases/chemistry
Stilbenes/chemistry
Sulfonic Acids/chemistry
Trichloroacetic Acid/chemistry

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10.1016/j.jim.2008.09.006

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title = "Rapid acidification and alkylation: redox analysis of the MHC class I pathway",

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.",

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author = "Powis, \{Simon J\} and Darren Nesbeth and Izabela Lenart and Helen Fussell and Tracey Lamb and Keith Gould and Antoniou, \{Antony N\}",

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doi = "10.1016/j.jim.2008.09.006",

language = "English",

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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

UR - https://www.scopus.com/pages/publications/57349127343

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 -

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

Abstract

Keywords

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