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A Sensitive Radioassay for Sulfhydryl Groups with Tetraethylthiuram Disulfide

From the ¹ From The Department of Physiological Chemistry, The Johns Hopkins University School of Medicine, Baltimore, Maryland 21205

A sensitive, specific, convenient radioassay for sulfhydryl groups with tetraethylthiuram disulfide that is uniformly labeled with radioactive sulfur is described. Interaction between reagent and thiol results in the formation of stoichiometric amounts of ³⁵S-diethyldithiocarbamate anion; the latter compound decomposes to labeled carbon disulfide at pH 4.0. The ³⁵S-carbon disulfide is allowed to diffuse from the reaction medium to alkaline piperidine, and its radioactivity, which is proportional to the sulfhydryl content of the sample, is evaluated. The sulfhydryl contents of five "native" and denatured proteins were determined; the results agreed with reported values. This nonspectrophotometric method has proved applicable for assay of the thiol content of as little as 10 µg of various proteins.

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