Estimation of protein sulfhydryl groups with 5,5'-(35S) dithio-bis(2-nitrobenzoate) |
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Authors: | P M Steinert B S Baliga H N Munro |
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Institution: | Physiological Chemistry Laboratories, Department of Nutrition and Food Science, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, USA |
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Abstract: | 35S-Labeled dithio-bis(2-nitrobenzoate) has been prepared and its usefulness for the measurement of protein sulfhydryl groups has been investigated. The technique is at least 20 times more sensitive than when dithio-bis(2-nitrobenzoate) is used colorimetrically. The adduct formed between the reagent and sulfhydryl groups is stable between pH 3–9 which should enable use of the reagent for the tagging of sulfhydryl-containing proteins in a wide variety of biochemical procedures. Removal of the adduct from a protein is achieved simply by treatment with a thiol reagent, with concomitant restoration of enzymic activity of the protein. The procedure has been successfully tested on alcohol dehydrogenase and mammalian ribosomes. |
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