Activation of a thioesterase specific for very-long-chain fatty acids by adrenergic agonists in perfused hearts.
We have recently described an acyl-CoA thioesterase specific for very-long-chain fatty acids, named ARTISt, that regulates steroidogenesis through the release of arachidonic acid in adrenal zona fasciculata cells. In this paper we demonstrate the presence of the protein as a 43 kDa band and its mRNA in cardiac tissue. The activity of the protein was measured using an heterologous cell-free assay in which it is recombined with adrenal microsomes and mitochondria to activate mitochondrial steroidogenesis. Isoproterenol and phenylephrine activate the enzyme in a dose-dependent manner (10(-10)-10(-6) M). Both propranolol (10(-5) M) and prazosin (10(-5) M) block the action of isoproterenol and phenylephrine respectively. Antipeptide antibodies against the serine lipase motif of the protein and the Cys residue present in the catalytic domain also block the activity of the protein. Taken together, our results confirm the presence of ARTISt in heart and provide evidence for a catecholamine-activated regulatory pathway of the enzyme in that tissue.
Neuman
I
I
Department of Biochemistry, School of Medicine, University of Buenos Aires, Paraguay 2155, 5 piso, 1121, Buenos Aires, Argentina.
Lisdero
C
C
Finkielstein
C
C
Maloberti
P
P
Mendez
C F
CF
Poderoso
J J
JJ
Podestá
E J
EJ
D000322:Adrenergic Agonists / Q000494:pharmacology*; D000818:Animals; D000906:Antibodies / Q000276:immunology; D017868:Cyclic AMP-Dependent Protein Kinases / Q000037:antagonists & inhibitors; D004305:Dose-Response Relationship, Drug; D004789:Enzyme Activation / Q000187:drug effects; D005227:Fatty Acids / Q000378:metabolism*; D006321:Heart / Q000187:drug effects*; D007545:Isoproterenol / Q000494:pharmacology; D009637:Masoprocol / Q000494:pharmacology; D024101:Mitochondrial Proteins; D009206:Myocardium / Q000201:enzymology; D010170:Palmitoyl-CoA Hydrolase / Q000378:metabolism; D010477:Perfusion; D010656:Phenylephrine / Q000494:pharmacology; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D017208:Rats, Wistar; D013869:Thiolester Hydrolases / Q000032:analysis / Q000276:immunology / Q000378:metabolism*
D000322:Adrenergic Agonists; D000906:Antibodies; D005227:Fatty Acids; D024101:Mitochondrial Proteins; D012333:RNA, Messenger; D010656:Phenylephrine; D009637:Masoprocol; D017868:Cyclic AMP-Dependent Protein Kinases; D013869:Thiolester Hydrolases; C546434:Acot2 protein, rat; D010170:Palmitoyl-CoA Hydrolase; D007545:Isoproterenol