Acyl-CoA ligases from rat brain microsomes: an immunochemical study.
Acyl-CoA ligase activities, solubilized from rat brain microsomes, were fractionated into three different peaks by hydroxyapatite chromatography. Based on physical and chemical properties, we suggested that peak A (pamitoyl-CoA ligase) and peak C (lignoceroyl-CoA ligase) were two different enzymes (A. Bhushan, R. P. Singh, and I. Singh (1986) Arch. Biochem. Biophys. 246, 374-380). We raised antibodies against purified liver microsomal palmitoyl-CoA ligase (EC 6.2.1.3) and examined the effect of this antibody on acyl-CoA ligase activities for palmitic, arachidonic and lignoceric acids in microsomal enzyme extract and different acyl-CoA ligase peaks from the hydroxyapatite column. In an enzyme activity assay system in microsomal extract, the antisera inhibited the palmitoyl-CoA ligase activity but had very little effect on the acyl-CoA ligase activities for arachidonic and lignoceric acids. This antisera inhibited the acyl-CoA ligase activities for these three fatty acids in peak A and had no effect on these activities in peak B or peak C. Western blot analysis demonstrated that antibody to liver microsomal palmitoyl-CoA ligase cross-reacted with only peak A (palmitoyl-CoA ligase), but not with peak B or peak C. This immunochemical study demonstrates that palmitoyl-CoA ligase does not share immunological determinants with acyl-CoA ligases in peaks B or C, thus demonstrating that palmitoyl-CoA ligase (peak A) is different from the arachidonoyl-CoA and lignoceroyl-CoA ligase activities in peaks B or C.
Singh
I
I
Department of Pediatrics, Medical University of South Carolina, Charleston 29425.
Bhushan
A
A
Relan
N K
NK
Hashimoto
T
T
D000818:Animals; D016718:Arachidonic Acid; D001095:Arachidonic Acids / Q000378:metabolism; D001921:Brain / Q000201:enzymology* / Q000648:ultrastructure; D003066:Coenzyme A Ligases / Q000378:metabolism*; D003429:Cross Reactions; D005227:Fatty Acids / Q000378:metabolism; D007150:Immunohistochemistry; D008861:Microsomes / Q000201:enzymology*; D051381:Rats; D012097:Repressor Proteins; D029701:Saccharomyces cerevisiae Proteins; D012995:Solubility; D000098859:Long-Chain-Fatty-Acid-CoA Ligase
D001095:Arachidonic Acids; D005227:Fatty Acids; D012097:Repressor Proteins; D029701:Saccharomyces cerevisiae Proteins; D016718:Arachidonic Acid; D003066:Coenzyme A Ligases; C044125:lignoceroyl-CoA ligase; C090799:FAA2 protein, S cerevisiae; D000098859:Long-Chain-Fatty-Acid-CoA Ligase; C010210:lignoceric acid