TY - JOUR
T1 - Cross-linking protein glutathionylation mediated by O2-arylated bis-diazeniumdiolate "double JS-K"
AU - Holland, Ryan J.
AU - MacIag, Anna E.
AU - Kumar, Varun
AU - Shi, Lei
AU - Saavedra, Joseph E.
AU - Prud'Homme, Robert K.
AU - Chakrapani, Harinath
AU - Keefer, Larry K.
N1 - Copyright:
Copyright 2013 Elsevier B.V., All rights reserved.
PY - 2012/12/17
Y1 - 2012/12/17
N2 - Attachment of glutathione (GSH) to cysteine residues in proteins (S-glutathionylation) is a reversible post-translational modification that can profoundly alter protein structure and function. Often serving in a protective role, for example, by temporarily saving protein thiols from irreversible oxidation and inactivation, glutathionylation can be identified and semiquantitatively assessed using anti-GSH antibodies, thought to be specific for recognition of the S-glutathionylation modification. Here, we describe an alternate mechanism of protein glutathionylation in which the sulfur atoms of the GSH and the protein's thiol group are covalently bound via a cross-linking agent, rather than through a disulfide bond. This form of thiol cross-linking has been shown to occur and has been confirmed by mass spectrometry at the solution chemistry level, as well as in experiments documenting the potent antiproliferative activity of the bis-diazeniumdiolate Double JS-K in H1703 cells in vitro and in vivo. The modification is recognized by the anti-GSH antibody as if it were authentic S-glutathionylation, requiring mass spectrometry to distinguish between them.
AB - Attachment of glutathione (GSH) to cysteine residues in proteins (S-glutathionylation) is a reversible post-translational modification that can profoundly alter protein structure and function. Often serving in a protective role, for example, by temporarily saving protein thiols from irreversible oxidation and inactivation, glutathionylation can be identified and semiquantitatively assessed using anti-GSH antibodies, thought to be specific for recognition of the S-glutathionylation modification. Here, we describe an alternate mechanism of protein glutathionylation in which the sulfur atoms of the GSH and the protein's thiol group are covalently bound via a cross-linking agent, rather than through a disulfide bond. This form of thiol cross-linking has been shown to occur and has been confirmed by mass spectrometry at the solution chemistry level, as well as in experiments documenting the potent antiproliferative activity of the bis-diazeniumdiolate Double JS-K in H1703 cells in vitro and in vivo. The modification is recognized by the anti-GSH antibody as if it were authentic S-glutathionylation, requiring mass spectrometry to distinguish between them.
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U2 - 10.1021/tx3003142
DO - 10.1021/tx3003142
M3 - Article
C2 - 23106594
AN - SCOPUS:84871249632
SN - 0893-228X
VL - 25
SP - 2670
EP - 2677
JO - Chemical Research in Toxicology
JF - Chemical Research in Toxicology
IS - 12
ER -