PMID: 3766964Aug 15, 1986Paper

A thin-gel isoelectric focusing method for quantitation of protein S-thiolation

Analytical Biochemistry
J A Thomas, D Beidler

Abstract

A thin-gel isoelectric focusing method has been developed for analysis of protein S-thiolation (formation of mixed disulfides with low molecular weight thiols). The method is rapid and it can be used with 3 to 5 micrograms of a pure protein, or 15 to 20 micrograms of tissue extract protein. It is possible to detect a modification of the protein sulfhydryl by either charged or uncharged thiols, and to determine the quantity of different S-thiolated protein species in a modified sample. The method was used to quantitate the amount of S-thiolation of phosphorylase b in a reaction with oxidized glutathione that produced four S-thiolated forms of the enzyme. The method was also used to detect S-thiolation of two proteins in a cardiac tissue extract treated with diamide. One of the protein bands was shown to be S-thiolated with both cysteine and glutathione, while the other band was S-thiolated only with glutathione.

References


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Citations

Jan 1, 1989·Archives of Dermatological Research·K KatsuumiY Sato
Dec 11, 1991·Biochemical Pharmacology·R B BirgeL A Smolin
Apr 22, 1987·Biochimica Et Biophysica Acta·M W Collison, J A Thomas
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Jul 16, 2010·Geriatrics & Gerontology International·Tosifusa TodaYoji Yoshimi
Jul 21, 1998·The Journal of Biological Chemistry·H H Wu, J Momand

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