S‐methanesulfonyl‐CoA: A thiol‐specific reagent for affinity labeling of short chain acyl‐CoA sites

Owens, Martha S.; Clements, Peter R.; Anderson, A.Duane; Barden, Roland E.

doi:10.1016/0014-5793(81)80124-x

Cited by 6 publications

(6 citation statements)

References 14 publications

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“…Other CoA analogues have been prepared by derivatization of the thiol group of CoA. These include S -methanesulfonyl-CoA, prepared by reaction of CoA with methanesulfonyl chloride . This compound rapidly inactivates succinate thiokinase and fatty acid synthase.…”

Section: E Other Thiol-modified Coa Derivativesmentioning

confidence: 99%

Coenzyme A Analogues and Derivatives: Synthesis and Applications as Mechanistic Probes of Coenzyme A Ester-Utilizing Enzymes

Mishra

Drueckhammer

2000

Chem. Rev.

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Section: E Other Thiol-modified Coa Derivativesmentioning

confidence: 99%

Coenzyme A Analogues and Derivatives: Synthesis and Applications as Mechanistic Probes of Coenzyme A Ester-Utilizing Enzymes

Mishra

Drueckhammer

2000

Chem. Rev.

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“…A thiosulfonate containing reagent, reported by Nishimura et al (1982), incorporated two CoA nucleotides and, due to steric considerations (i.e., a potential requirement for two nucleotide binding pockets), its general utility with many acyl-CoA utilizing enzymes might be expected to be limited. S-Methanesulfonyl-CoA, prepared by reaction of methanesulfonyl chloride with CoASH (Owens et al, 1981), proved effective as an inactivator of succinate thiokinase and β-hydroxyacyl-CoA dehydrogenase. However, it was also reported to function as a simple group specific reagent, presumably due to the poor binding specificity afforded by the methyl group.…”

Section: Methodsmentioning

confidence: 99%

Inactivation of 3-Hydroxy-3-methylglutaryl-CoA Synthase and Other Acyl-CoA-Utilizing Enzymes by 3-Oxobutylsulfoxyl-CoA

1997

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3-Oxobutylsulfoxyl-CoA has been produced by oxidation of S-3-oxobutyl-CoA, the thioether analog of acetoacetyl-CoA. Avian hydroxymethylglutaryl-CoA (HMG-CoA) synthase is inactivated by oxobutylsulfoxyl-CoA in a time-dependent fashion. Protection against inactivation is afforded by the substrate, acetyl-CoA, suggesting that inactivation involves modification of the enzyme's active site. Pretreatment of HMG-CoA synthase with the inactivator blocks the enzyme's ability to form Michaelis and acetyl-S-enzyme intermediates, supporting the hypothesis that modification is active-site directed. Incubation of enzyme with oxobutylsulfoxyl-[32P]CoA followed by precipitation with trichloroacetic acid indicates that inactivation correlates with stoichiometric formation of a covalent adduct between enzyme and a portion of the inactivator that includes the CoA nucleotide. The observation of reagent partitioning suggests that HMG-CoA synthase catalyzes conversion of oxobutylsulfoxyl-CoA into a reactive species that modifies the protein. Treatment of inactivated enzyme with DTT or other mercaptans restores enzyme activity and reverses the covalent modification with release of CoASH. Oxobutylsulfoxyl-CoA inactivates beta-ketothiolase and HMG-CoA lyase in a process that is also reversed by DTT. These three enzymes all contain active site cysteines, suggesting that inactivation results from disulfide formation between a cysteine and the CoA moiety of the inhibitor. The data are consistent with the hypothesis that enzymatic cleavage of oxobutylsulfoxyl-CoA results in the transient formation of a sulfenic acid derivative of CoA which subsequently reacts to form a stable disulfide linkage to protein.

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“…2-Bromopalmitoyl-CoA was synthesized from 2-bromopalmitic acid via 2-bromopalmitoyl chloride as described by Chase & Tubbs (1972). For methanesulphonyl-CoA, the method of Owens et al (1981) was modified in that 10mg of CoA were made to react at 0°C with 101 of methanesulphonyl chloride in 1 M-KHCO3, pH 8.0, with vigorous shaking for 30s. The product, after diethyl ether extractions, was not further purified.…”

Section: Chemical Synthesismentioning

confidence: 99%

“…Similar experiments were not attempted for mixtures of bromopalmitoyl-CoA and malonyl-CoA or bromoacetyl-CoA, because inhibition by bromopalmitoyl-CoA was not reversible and because [free bromopalmitoyl-CoA] may not increase linearly with [total bromopalmitoyl-CoA] in the presence of albumin. S-Methanesulphonyl-CoA is an active-sitedirected inhibitor of succinic thiokinase and 3hydroxyacyl-CoA dehydrogenase (Owens et al, 1981). It has been suggested that, because of the high specificity of the thiolsulphonate group for thiols, S-methanesulphonyl-CoA could be used to detect the presence of a reactive thiol in an acyl-CoA-binding site (Owens et al, 1981).…”

Section: Inhibitors and Other Additionsmentioning

confidence: 99%

“…S-Methanesulphonyl-CoA is an active-sitedirected inhibitor of succinic thiokinase and 3hydroxyacyl-CoA dehydrogenase (Owens et al, 1981). It has been suggested that, because of the high specificity of the thiolsulphonate group for thiols, S-methanesulphonyl-CoA could be used to detect the presence of a reactive thiol in an acyl-CoA-binding site (Owens et al, 1981). S-Methanesulphonyl-CoA was found to inhibit liver CPT, (Fig.…”

Section: Inhibitors and Other Additionsmentioning

confidence: 99%

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Effects of dl-2-bromopalmitoyl-CoA and bromoacetyl-CoA in rat liver and heart mitochondria. Inhibition of carnitine palmitoyltransferase and displacement of [14C]malonyl-CoA from mitochondrial binding sites

Edwards¹,

Bird²,

Saggerson³

1985

Biochemical Journal

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The overt form of carnitine palmitoyltransferase (CPT1) in rat liver and heart mitochondria was inhibited by DL-2-bromopalmitoyl-CoA and bromoacetyl-CoA. S-Methanesulphonyl-CoA inhibited liver CPT1. The inhibitory potency of DL-2-bromopalmitoyl-CoA was 17 times greater with liver than with heart CPT1. Inhibition of CPT1 by DL-2-bromopalmitoyl-CoA was unaffected by 5,5'-dithiobis-(2-nitrobenzoic acid) or (in liver) by starvation. In experiments in which DL-2-bromopalmitoyl-CoA displaced [14C]malonyl-CoA bound to liver mitochondria, the KD (competing) was 25 times the IC50 for inhibition of CPT1 providing evidence that the malonyl-CoA-binding site is unlikely to be the same as the acyl-CoA substrate site. Bromoacetyl-CoA inhibition of CPT1 was more potent in heart than in liver mitochondria and was diminished by 5,5'-dithiobis-(2-nitrobenzoic acid) or (in liver) by starvation. Bromoacetyl-CoA displaced bound [14C]malonyl-CoA from heart and liver mitochondria. In heart mitochondria this displacement was competitive with malonyl-CoA and was considerably facilitated by L-carnitine. In liver mitochondria this synergism between carnitine and bromoacetyl-CoA was not observed. It is suggested that bromoacetyl-CoA interacts with the malonyl-CoA-binding site of CPT1. L-Carnitine also facilitated the displacement by DL-2-bromopalmitoyl-CoA of [14C]malonyl-CoA from heart, but not from liver, mitochondria. DL-2-Bromopalmitoyl-CoA and bromoacetyl-CoA also inhibited overt carnitine octanoyl-transferase in liver and heart mitochondria. These findings are discussed in relation to inter-tissue differences in (a) the response of CPT1 activity to various inhibitors and (b) the relationship between high-affinity malonyl-CoA-binding sites and those sites for binding of L-carnitine and acyl-CoA substrates.

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S‐methanesulfonyl‐CoA: A thiol‐specific reagent for affinity labeling of short chain acyl‐CoA sites

Cited by 6 publications

References 14 publications

Coenzyme A Analogues and Derivatives: Synthesis and Applications as Mechanistic Probes of Coenzyme A Ester-Utilizing Enzymes

Coenzyme A Analogues and Derivatives: Synthesis and Applications as Mechanistic Probes of Coenzyme A Ester-Utilizing Enzymes

Inactivation of 3-Hydroxy-3-methylglutaryl-CoA Synthase and Other Acyl-CoA-Utilizing Enzymes by 3-Oxobutylsulfoxyl-CoA

Effects of dl-2-bromopalmitoyl-CoA and bromoacetyl-CoA in rat liver and heart mitochondria. Inhibition of carnitine palmitoyltransferase and displacement of [14C]malonyl-CoA from mitochondrial binding sites

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