Abstract
Fatty acid ethyl ester synthase-III metabolizes both ethanol and carcinogens. Structure-function studies of the enzyme have not been performed in relation to site specific mutagenesis. In this study, three residues (Gly 32, Cys 39 and His 72) have been mutated to observe their role in enzyme activity. Gly to Gln, Cys to Trp and His to Ser mutations did not affect fatty acid ethyl ester synthase activity, but His to Ser mutant had less than 9% of control glutathione S-transferase activity. The apparent loss of transferase activity reflected a 28 fold weaker binding constant for glutathione. Thus, this study indicates that Gly and Cys may not be important for synthase or transferase activities however, histidine may play a role in glutathione binding, but it is not an essential catalytic residue of glutathione S-transferase or for fatty acid ethyl ester synthase activity.
Similar content being viewed by others
References
Bora PS, Spilburg CA, Lange LG: Metabolism of ethanol and carcinogens by glutathione transferases. Proc Natl Acad Sci USA 86: 4470–4473, 1989
Bora PS, Spilburg CA, Lange LG: Identification of a satelite fatty acid ethyl ester synthase from human myocardium as a glutathione Stransferase. J Clin Invest 84: 1942–1946, 1989
Bora PS, Spilburg CA, Lange LG: Purification to homogeneity and characterization of major fatty acid ethyl ester synthase from human myocardium. FEBS Lett 258: 236–239, 1989
Bora PS, Lange LG: In: R.R. Watson, (ed). Alcohol and Drug Abuse Reviews. Humana Press, Inc. Clifton, NJ. Vol. 2 1991: pp. 241–257
Bora PS, Lange LG: Molecular mechanism of ethanol metabolism by human brain to fatty acid ethyl esters (review). Alcoholism: Clin and Exp Res 17: 28–30, 1993
Bora PS, Lange LG: Homogenous synthase I from human myocardium is a glutathione S-transferase. Ann NY Acad Sci 625: 827–829, 1991
Bora PS, Bora NS, Wu X, Lange LG: Molecular cloning, sequencing, and expression of human myocardial fatty acid ethyl ester synthase-III cDNA. J Biol Chem 266: 16774–16777, 1991
Kunkel TA, Roberts JD, Zakour RA: Rapid and efficient site-specific mutagenesis without pheotypic selection. Meth Enzymol 154: 367–382, 1987
Yanisch-Perron C, Vieira J, Messing T: Improved M13 phage clong vectors and host strains: nucleotide sequences of the M13 mp18 and puc19 vectors. Gene (amst) 33: 103–119, 1985
Sanger F, Nicklen S, Coulson AR: DNA sequencing with chainterminating inhibitors. Proc Natl Acad Sci USA 74: 5463–5467, 1977
Sussman DT, Milman G: Short-term high efficiency expression of transfected DNA. Mol Cell Biol 4: 1641–1645, 1984
Laemmli UK: Clearage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685, 1970
Reinemer P, Dirr HW, Ladenstein R, Schaffer J, Gallay O, Huber R: The three dimensional structure of class pi glutathione S-transferase in complex with glutattione sulfonate at 2.3A resolution. EMBO J 10: 1997–2005, 1991
Bora PS, Miller DD, Chaitman BR, Guruge BL: Myocardial cell damage by fatty acid ethyl esters. J Cardiovas Pharm 27: 1–6, 1997
Bora PS, Guruge BL, Miller DD, Chaitman BR, Ruyle MS: Purification and characterization of human heart fatty acid ethyl ester synthase/carboxylesterase. J Mol Cell Cardiol 28: 2027–2032, 1996
Ballester MR, Marti V, Guardia J, Riera JM: Spectrum of alcoholinduced myocardial necrosis detected by IIIIn-monoclonal antimyosin antibodies. J Am Coll Cardiol 25: 200–206, 1997
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Bora, P.S., Miller, D.D. & Chaitman, B.R. Mutagenesis and characterization of specific residues in fatty acid ethyl ester synthase: A gene for alcohol-induced cardiomyopathy. Mol Cell Biochem 180, 111–115 (1998). https://doi.org/10.1023/A:1006899208831
Issue Date:
DOI: https://doi.org/10.1023/A:1006899208831