Complementary DNAs encoding rat long-chain acyl-CoA synthetase have been isolated. The cDNAs were identified using synthetic oligonucleotide probes based on partial amino acid sequences of lysyl endopeptidase peptides of the purified enzyme. Rat long-chain acyl-CoA synthetase is predicted to contain 699 amino acid residues and to have a calculated molecular weight of 78,177. Significant sequenc...

The enzymes catalysing the initial stage of the 8-oxidation of fatty acids, the acyl-CoA synthetases, have been classified into four groups based on specificity. These are: the short-chain (acetyl-CoA synthetase; EC 6.2.1 .l), medium-chain (butyryl-CoA synthetase; EC 6.2.1.2) and the long-chain fatty acyl-CoA synthetase (acyl-CoA synthetase; EC 6.2.1.3), which are ATP-dependent and follow the r...

The kinetics of activation of saturated fatty acids by long chain acyl-CoA synthetase from rat liver microsomes has been studied with a method of selective extraction of free fatty acids based on the insolubility of acyl-CoA in diethyl ether. Saturated fatty acids with a chain length ranging from Cl* to Co were assayed at concentrations varying from 0.5 to 10 PM. Under these conditions, Vm,, is...

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 sol...

The possibility of a regulatory role of long-chain acyl-CoAs was studied in a system synthesizing palmitoyl-CoA and stearoyl-CoA in vitro. This system contained acetyl-CoA carboxylase (yeast), fatty acid synthetase (yeast) and lipid membranes as acceptors for the long-chain acyl-CoA compounds. In this system acetyl-CoA carboxylase is controlled by long-chain acyl-CoAs, whereas this is not true ...

Fatty acyl-CoA synthetase, the first enzyme of the beta-oxidation pathway, has been proposed to be involved in long chain fatty acid translocation across the plasma membrane of prokaryotic and eukaryotic cells. To test this proposal, we used an in vitro system consisting of Escherichia coli inner (plasma) membrane vesicles containing differing amounts of trapped fatty acyl-CoA synthetase and it...

Fatty acyl-CoA synthetase (fatty acid: CoA ligase, AMP-forming; (EC 6.2.1.3)) catalyzes the formation of fatty acyl-CoA by a two-step process that proceeds through the hydrolysis of pyrophosphate. Fatty acyl-CoA represents bioactive compounds that are involved in protein transport, enzyme activation, protein acylation, cell signaling, and transcriptional control in addition to serving as substr...

It is shown that acyl-CoA binding protein (ACBP), in contrast with fatty acid binding protein (FABP), stimulates the synthesis of long-chain acyl-CoA esters by mitochondria. ACBP effectively opposes the product feedback inhibition of the long-chain acyl-CoA synthetase by sequestration of the synthesized acyl-CoA esters. Feedback inhibition of microsomal long-chain acyl-CoA synthesis could not b...

Triglyceride and phospholipids are the main constituents of glycerolipids in eucaryotic cells. They are synthesized via a common precursor, phosphatidic acid, although their physiological roles are entirely different. Despite the biological importance of these two lipid classes, little is known about the control mechanism involved in their synthesis. We have shown previously that there are two ...

Recently we found that firefly luciferase is a bifunctional enzyme, catalyzing not only the luminescence reaction but also long-chain fatty acyl-CoA synthesis. Further, the gene product of CG6178 (CG6178), an ortholog of firefly luciferase in Drosophila melanogaster, was found to be a long-chain fatty acyl-CoA synthetase and dose not function as a luciferase. We investigated the substrate speci...