Methoxycarbonyl-CoA Disulfide: An Active Site=Directed Sulfhydryl Reagent

Louis B. Hersh; Daniel J. Smith

doi:10.1016/S0076-6879(81)72047-0

Methoxycarbonyl-CoA Disulfide: An Active Site=Directed Sulfhydryl Reagent

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Abstract

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.

Original language	English
Pages (from-to)	584-587
Number of pages	4
Journal	Methods in Enzymology
Volume	72
Issue number	C
DOIs	https://doi.org/10.1016/S0076-6879(81)72047-0
State	Published - Jan 1 1981

Bibliographical note

Funding 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

Biochemistry
Molecular Biology

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10.1016/S0076-6879(81)72047-0

Cite this

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title = "Methoxycarbonyl-CoA Disulfide: An Active Site=Directed Sulfhydryl Reagent",

abstract = "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.",

author = "Hersh, {Louis B.} and Smith, {Daniel J.}",

note = "Funding 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).",

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N2 - 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.

AB - 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.

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Methoxycarbonyl-CoA Disulfide: An Active Site=Directed Sulfhydryl Reagent

Abstract

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