Saccharomyces cerevisiae contains at least five acyl-CoA synthetases (fatty acid activation proteins, or Faaps). Four FAA genes have been recovered to date. Recent genetic studies indicate that Faa1p and Faa4p are involved in the activation of imported fatty acids, while Faa2p activates endogenous pools of fatty acids. We have now purified Faa4p from S. cerevisiae and compared its fatty acid substrate specificity in vitro with the specificities of purified Faa1p, Faa2p, and Faa3p. Among C8-C18 saturated fatty acids, Faa4p and Faa1p both prefer C14:0. Surveys of C14 fatty acids with single cis-double bonds at C2-C12 indicated that Faa4p and Faa1p prefer Z9-tetradecenoic acid, although Faa4p's preference is much greater and also evident in C16 and C18 fatty acids. Faa4p's selectivity for fatty acids with a C9-C10 cis-double bond is a feature it shares with Faa3p and is notable since in yeast Ole1p, a microsomal cis- desaturase, accounts for de novo production of monoenoic acyl-CoAs from saturated acyl-CoA substrates. Faa4p has no detectable acyl-CoA synthetase activity when incubated with tetradecenoic acids having a trans-double bond at C2-3, C4-5, C5-6, C6-7, C7-8, or C9-10. Faa3p can only use E9-tetradecenoic acid as a substrate, while E4-, E6- and E9-tetradecenoic acids can be used by Faa1p and Faa2p. E2-tetradecenoic acid is an Faap inhibitor, with Faa2p exhibiting the greatest sensitivity (IC = 2.6 ± 0.2 µM). Triacsin C (1-hydroxy3-(E,E,E,2`,4`,7`-undecatrienylidine)-1,2,3-triazene) has trans-double bonds at positions that correspond to those in E2-, E5-, and E7-tetradecenoic acids. This compound is a potent inhibitor of Faa2p (K = 15 ± 1 nM; competitive with fatty acid), less potent against Faa4p (K = 2 µM), and not active against Faa1p or Faa3p (IC > 500 µM). Analysis of an n-tetradecanal plus a series of oximes (tridecanal oxime, 1-azadeca-1,3,5-trienol, and 1-azaundeca-1,3,5-trienol) indicated that the combination of an azenol moiety (R-CH&cjs0808;N-OH) plus adjacent unsaturation are critical for triacsin C's selective inhibition of Faa2p. Triacsin C and oxime derivatives appear to be very useful for defining differences in molecular recognition among S. cerevisiae acyl-CoA synthetases. The >25,000-fold range in the inhibitory effects of triacsin C on these four Faaps suggests that it may be possible to develop other selective inhibitors of eukaryotic acyl-CoA synthetases.

CiteULike

Complore

Connotea

Del.icio.us

Digg

Reddit

Technorati

This article has been cited by other articles:

				M. S. Han, S. Y. Park, K. Shinzawa, S. Kim, K. W. Chung, J.-H. Lee, C. H. Kwon, K.-W. Lee, J.-H. Lee, C. K. Park, et al. Lysophosphatidylcholine as a death effector in the lipoapoptosis of hepatocytes J. Lipid Res., January 1, 2008; 49(1): 84 - 97. [Abstract] [Full Text] [PDF]

				M. T Weis, J. L Crumley, L. H Young, and J. N Stallone Inhibiting long chain fatty Acyl CoA synthetase increases basal and agonist-stimulated NO synthesis in endothelium Cardiovasc Res, August 1, 2004; 63(2): 338 - 346. [Abstract] [Full Text] [PDF]

				Z. Zou, F. Tong, N. J. Fargeman, C. Borsting, P. N. Black, and C. C. DiRusso Vectorial Acylation in Saccharomyces cerevisiae. Fat1p AND FATTY ACYL-CoA SYNTHETASE ARE INTERACTING COMPONENTS OF A FATTY ACID IMPORT COMPLEX J. Biol. Chem., April 25, 2003; 278(18): 16414 - 16422. [Abstract] [Full Text] [PDF]

				Y.-M. Mu, T. Yanase, Y. Nishi, A. Tanaka, M. Saito, C.-H. Jin, C. Mukasa, T. Okabe, M. Nomura, K. Goto, et al. Saturated FFAs, Palmitic Acid and Stearic Acid, Induce Apoptosis in Human Granulosa Cells Endocrinology, August 1, 2001; 142(8): 3590 - 3597. [Abstract] [Full Text] [PDF]

				D. M. Muoio, T. M. Lewin, P. Wiedmer, and R. A. Coleman Acyl-CoAs are functionally channeled in liver: potential role of acyl-CoA synthetase Am J Physiol Endocrinol Metab, December 1, 2000; 279(6): E1366 - E1373. [Abstract] [Full Text] [PDF]

				Y. Hernando, A. T. Carter, A. Parr, B. Hove-Jensen, and M. Schweizer Genetic Analysis and Enzyme Activity Suggest the Existence of More Than One Minimal Functional Unit Capable of Synthesizing Phosphoribosyl Pyrophosphate in Saccharomyces cerevisiae J. Biol. Chem., April 30, 1999; 274(18): 12480 - 12487. [Abstract] [Full Text] [PDF]

				K. Ashrafi, T. A. Farazi, and J. I. Gordon A Role for Saccharomyces cerevisiae Fatty Acid Activation Protein 4 in Regulating Protein N-Myristoylation during Entry into Stationary Phase J. Biol. Chem., October 2, 1998; 273(40): 25864 - 25874. [Abstract] [Full Text] [PDF]

				R. A. Igal and R. A. Coleman Acylglycerol Recycling from Triacylglycerol to Phospholipid, Not Lipase Activity, Is Defective in Neutral Lipid Storage Disease Fibroblasts J. Biol. Chem., July 12, 1996; 271(28): 16644 - 16651. [Abstract] [Full Text] [PDF]

				T. M. Lewin, J.-H. Kim, D. A. Granger, J. E. Vance, and R. A. Coleman Acyl-CoA Synthetase Isoforms 1, 4, and 5 Are Present in Different Subcellular Membranes in Rat Liver and Can Be Inhibited Independently J. Biol. Chem., June 29, 2001; 276(27): 24674 - 24679. [Abstract] [Full Text] [PDF]