S-thiolation of HSP27 regulates its multimeric aggregate size independently of phosphorylation

Eaton, P., Fuller, W. and Shattock, M. J. (2002) S-thiolation of HSP27 regulates its multimeric aggregate size independently of phosphorylation. Journal of Biological Chemistry, 277(24), pp. 21189-21196. (doi:10.1074/jbc.M200591200) (PMID:11925435)

Full text not currently available from Enlighten.

Abstract

HSP27 exists as large aggregates that breakdown after phosphorylation. We show rat cardiac HSP27 isS-thiolated during oxidant stress, and this modification,without phosphorylation, disaggregates multimeric HSP27. Biotinylated cysteine acts as a probe for thiolated proteins, which are detected using non-reducing Western blots probed with streptavidin-horseradish peroxidase. Controls show a low level of S-thiolation, which is increased 3.6-fold during post-ischemic reperfusion. S-Thiolated proteins were purified using streptavidin-agarose, and Western immunoblotting showed HSP27 was present. We increased protein S-thiolation 10-fold with 10 μM H2O2 with or without a kinase inhibitor mixture (staurosporine, genistein, bisindolylmaleimide, SB203580, and PD98059). H2O2 alone induced the phosphorylation of HSP27 Ser-86 and Ser-45/Ser-59 of its homologue αB crystallin. However, kinase inhibition reduced phosphorylation of these sites below basal. Despite effective kinase inhibition, H2O2 still disaggregated HSP27, but not αB crystallin. This is consistent with the lack of an S-thiolation site on αB crystallin. Thus, we have demonstrated a novel mechanism of HSP27 multimeric size regulation. S-Thiolation must occur at Cys-141, the only cysteine in rat HSP27.

Item Type:Articles
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:Fuller, Dr Will
Authors: Eaton, P., Fuller, W., and Shattock, M. J.
College/School:College of Medical Veterinary and Life Sciences > Institute of Cardiovascular and Medical Sciences
Journal Name:Journal of Biological Chemistry
Publisher:American Society for Biochemistry and Molecular Biology, Inc.
ISSN:0021-9258
ISSN (Online):1083-351X

University Staff: Request a correction | Enlighten Editors: Update this record