title: Molecular and biochemical characterization of rat gamma-trimethylaminobutyraldehyde dehydrogenase and evidence for the involvement of human aldehyde dehydrogenase 9 in carnitine biosynthesis.
abstract: The penultimate step in carnitine biosynthesis is mediated by gamma-trimethylaminobutyraldehyde dehydrogenase (EC 1.2.1.47), a cytosolic NAD(+)-dependent aldehyde dehydrogenase that converts gamma-trimethylaminobutyraldehyde into gamma-butyrobetaine. This enzyme was purified from rat liver, and two internal peptide fragments were sequenced by Edman degradation. The peptide sequences were used to search the Expressed Sequence Tag data base, which led to the identification of a rat cDNA containing an open reading frame of 1485 base pairs encoding a polypeptide of 494 amino acids with a calculated molecular mass of 55 kDa. Expression of the coding sequence in Escherichia coli confirmed that the cDNA encodes gamma-trimethylaminobutyraldehyde dehydrogenase. The previously identified human aldehyde dehydrogenase 9 (EC 1.2.1.19) has 92% identity with rat trimethylaminobutyraldehyde dehydrogenase and has been reported to convert substrates that resemble gamma-trimethylaminobutyraldehyde. When aldehyde dehydrogenase 9 was expressed in E. coli, it exhibited high trimethylaminobutyraldehyde dehydrogenase activity. Furthermore, comparison of the enzymatic characteristics of the heterologously expressed human and rat dehydrogenases with those of purified rat liver trimethylaminobutyraldehyde dehydrogenase revealed that the three enzymes have highly similar substrate specificities. In addition, the highest V(max)/K(m) values were obtained with gamma-trimethylaminobutyraldehyde as substrate. This indicates that human aldehyde dehydrogenase 9 is the gamma-trimethylaminobutyraldehyde dehydrogenase, which functions in carnitine biosynthesis.
authors.affiliation: Laboratory for Genetic Metabolic Diseases, Departments of Clinical Chemistry and Pediatrics, Emma Children's Hospital, Academic Medical Center, University of Amsterdam, P. O. Box 22700, 1100 DE Amsterdam, The Netherlands.
authors.forename: F M
authors.initials: FM
authors.lastname: Vaz
authors.forename: S W
authors.initials: SW
authors.lastname: Fouchier
authors.forename: R
authors.initials: R
authors.lastname: Ofman
authors.forename: M
authors.initials: M
authors.lastname: Sommer
authors.forename: R J
authors.initials: RJ
authors.lastname: Wanders
mesh_terms: D018528:ATP-Binding Cassette Transporters; D000444:Aldehyde Dehydrogenase / Q000235:genetics / Q000302:isolation & purification / Q000502:physiology*; D000447:Aldehydes / Q000378:metabolism*; D000595:Amino Acid Sequence; D000818:Animals; D002331:Carnitine / Q000096:biosynthesis*; D002352:Carrier Proteins / Q000096:biosynthesis; D018076:DNA, Complementary / Q000302:isolation & purification; D004926:Escherichia coli / Q000235:genetics; D029968:Escherichia coli Proteins; D006801:Humans; D008099:Liver / Q000201:enzymology; D058619:Maltose-Binding Proteins; D008969:Molecular Sequence Data; D009004:Monosaccharide Transport Proteins; D016366:Open Reading Frames; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D013379:Substrate Specificity
chemical_list: D018528:ATP-Binding Cassette Transporters; D000447:Aldehydes; D002352:Carrier Proteins; D018076:DNA, Complementary; D029968:Escherichia coli Proteins; D058619:Maltose-Binding Proteins; D009004:Monosaccharide Transport Proteins; D011993:Recombinant Fusion Proteins; C461599:maltose transport system, E coli; C029041:gamma-trimethylaminobutyraldehyde; D000444:Aldehyde Dehydrogenase; D002331:Carnitine