Purification and characterization of human heart fatty acid ethyl eater synthase/carboxylesterase

Puran S. Bora, Bandula L. Guruge, Donald D Miller, Bernard R. Chaitman, Matthew S. Ruyle

Research output: Contribution to journalArticle

26 Citations (Scopus)

Abstract

Fatty acid ethyl ester synthase metabolizes ethanol non-oxidatively in those extrahepatic organs most commonly damaged by alcohol abuse. This study was designed to purify human myocardial fatty acid ethyl ester synthase (FAEES)/carboxylesterase from human heart. The enzyme was purified to homogeneity after chromatography over DEAE-cellulose, Sephadex G-100 and hydroxylapatite. The homogenous enzyme, 62 kDa, has both synthase and carboxylesterase activities. The N-terminal amino acid sequence of the first 17 residues of the purified enzymes were 88% homologous to that of the carboxylesterase from rat liver and adipose tissue. Antibody was raised against pure synthase/carboxylesterase, cross-reacted with human cytosolic and microsomal fractions. With a constant oleic acid concentration of 0.25 mM, a calculated apparent K(m) and V(max) for ethanol were 0.30 M and 3700 nmol/mg protein/h., respectively. With constant ethanol concentrations of 1.2 M, the activity increased with the concentration of oleic acid to 0.17 mM, plateau to 0.25 mM. Because synthase/carboxylesterase esterifies free fatty acids with ethanol to produce its esters with potentially toxic effects, it may now be feasible to establish a link between alcohol consumption and end-organ damage.

Original languageEnglish (US)
Pages (from-to)2027-2032
Number of pages6
JournalJournal of molecular and cellular cardiology
Volume28
Issue number9
DOIs
StatePublished - Jan 1 1996
Externally publishedYes

Fingerprint

Carboxylesterase
Fatty Acids
Ethanol
Oleic Acid
Enzymes
DEAE-Dextran
Poisons
Durapatite
Nonesterified Fatty Acids
Alcohol Drinking
Alcoholism
Adipose Tissue
Chromatography
Amino Acid Sequence
Esters
Antibodies
Liver
Proteins

Keywords

  • Alcohol
  • Carboxylesterase
  • Enzyme
  • Fatty acid ethyl ester synthase
  • Myocardium

ASJC Scopus subject areas

  • Molecular Biology
  • Cardiology and Cardiovascular Medicine

Cite this

Purification and characterization of human heart fatty acid ethyl eater synthase/carboxylesterase. / Bora, Puran S.; Guruge, Bandula L.; Miller, Donald D; Chaitman, Bernard R.; Ruyle, Matthew S.

In: Journal of molecular and cellular cardiology, Vol. 28, No. 9, 01.01.1996, p. 2027-2032.

Research output: Contribution to journalArticle

Bora, Puran S. ; Guruge, Bandula L. ; Miller, Donald D ; Chaitman, Bernard R. ; Ruyle, Matthew S. / Purification and characterization of human heart fatty acid ethyl eater synthase/carboxylesterase. In: Journal of molecular and cellular cardiology. 1996 ; Vol. 28, No. 9. pp. 2027-2032.
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