Arachidonyl-CoA synthetase. Separation from nonspecific acyl-CoA synthetase and distribution in various cells and tissues

Michael Laposata, E. L. Reich, P. W. Majerus

Research output: Contribution to journalArticle

80 Citations (Scopus)

Abstract

Arachidonoyl-CoA synthetase was solubilized from a particulate fraction of calf brain and human platelets using 1% Nonidet P-40 and 10 mM EDTA. Arachidonoyl-CoA synthetase from both preparations was separated from nonspecific (long chain) acyl-CoA synthetase (EC 6.2.1.3) by chromatography on hydroxylapatite. To further substantiate that the two acyl-CoA synthetases are distinct proteins, we solubilized enzyme from a mutant cell line lacking arachidonoyl-CoA synthetase and from the parent cell line from which it was derived. These preparations were chromatographed on hydroxylapatite, and the mutant showed an absence of the peak identified as arachidonoyl-CoA synthetase in the parent while retaining the peak of nonspecific acyl-CoA synthetase activity. We have also determined the levels of arachidonoyl and nonspecific acyl-CoA synthetase in 13 different human cells and tissues. Arachidonoyl-CoA synthetase is widely distributed and is present in significantly lower concentrations than nonspecific acyl-CoA synthetase only in adipose tissue and liver.

Original languageEnglish (US)
Pages (from-to)11016-11020
Number of pages5
JournalJournal of Biological Chemistry
Volume260
Issue number20
StatePublished - 1985
Externally publishedYes

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Coenzyme A Ligases
Ligases
Tissue
Cells
Durapatite
Cell Line
Acyl Coenzyme A
Platelets
Chromatography
Edetic Acid
Liver
Adipose Tissue
Brain
Blood Platelets
arachidonyl-coenzyme A
arachidonate - CoA ligase
Enzymes
Proteins

ASJC Scopus subject areas

  • Biochemistry

Cite this

Arachidonyl-CoA synthetase. Separation from nonspecific acyl-CoA synthetase and distribution in various cells and tissues. / Laposata, Michael; Reich, E. L.; Majerus, P. W.

In: Journal of Biological Chemistry, Vol. 260, No. 20, 1985, p. 11016-11020.

Research output: Contribution to journalArticle

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AB - Arachidonoyl-CoA synthetase was solubilized from a particulate fraction of calf brain and human platelets using 1% Nonidet P-40 and 10 mM EDTA. Arachidonoyl-CoA synthetase from both preparations was separated from nonspecific (long chain) acyl-CoA synthetase (EC 6.2.1.3) by chromatography on hydroxylapatite. To further substantiate that the two acyl-CoA synthetases are distinct proteins, we solubilized enzyme from a mutant cell line lacking arachidonoyl-CoA synthetase and from the parent cell line from which it was derived. These preparations were chromatographed on hydroxylapatite, and the mutant showed an absence of the peak identified as arachidonoyl-CoA synthetase in the parent while retaining the peak of nonspecific acyl-CoA synthetase activity. We have also determined the levels of arachidonoyl and nonspecific acyl-CoA synthetase in 13 different human cells and tissues. Arachidonoyl-CoA synthetase is widely distributed and is present in significantly lower concentrations than nonspecific acyl-CoA synthetase only in adipose tissue and liver.

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