This chapter discusses the preparation and effects of methoxycarbonyl-CoA disulfide. The relative ease with which protein sulfhydryl groups reacted with a variety of compounds has led to the preparation of a large number of sulfhydryl reagents. These reagents have been classified according to the groups that are delivered, including blocking and labeling groups, reporter groups, cross-linking groups, and afffinity-abeling groups. Methoxycarbonyl-CoA disulfide is prepared by the reaction of free CoA with methoxycarbonylsulfenyi chloride. The latter compound is prepared in a two-step synthesis starting with trichloromethanesulfenyl chloride. Trichloromethanesulfenyl chloride is added to 211 g of concentrated sulfuric acid and 18 g of water. The mixture is stirred at 45–50° in a well-vented hood until hydrogen chloride evolution ceases. The methyl group of methoxycarbonyl-CoA disulfide can be replaced with a variety of alkyl groups by substituting different alcohols for methanol in the reaction with chlorocarbonylsulfenyl chloride. Radiolabeled reagent can be prepared by using 3H-labeled CoA. The reaction of methoxycarbonyl-CoA disulfide with choline acetyltransferase is also discussed.
|Number of pages||4|
|Journal||Methods in Enzymology|
|State||Published - Jan 1 1981|
Bibliographical noteFunding Information:
i This work was supported in part by Grant AM 13443 from The National Institutes of Health, Grant 1-391 from the Robert A. Welch Foundation, and Grant 10850-G4 from The Petroleum Research Foundation. la G. Kenyon and T. C. Bruice, this series, Vol. 47, p. 407. 2 D. J. Smith and G. L. Kenyon, J. Biol. Chem. 249, 3317 (1974). 3 D. J. Smith, E. T. Maggio, and G. L. Kenyon, Biochemistry 14, 766 (1975). 4 G. Zumach and E. Kuhle, Angew Chem., Int. Ed. Engl. 9, 54 (1970).
ASJC Scopus subject areas
- Molecular Biology