title: Purification, characterization, and mass spectrometric sequencing of a medium chain acyl-CoA synthetase from mouse liver mitochondria and comparisons with the homologues of rat and bovine.
abstract: Medium chain acyl-CoA synthetases catalyze the first reaction of amino acid conjugation of many xenobiotic carboxylic acids and fatty acid metabolism. This paper reports studies on purification, characterization, and the partial amino acid sequence of mouse liver enzyme. The medium chain acyl-CoA synthetase was isolated from mouse liver mitochondria. The purified enzyme catalyzes this reaction not only for straight medium chain fatty acids but also for aromatic and arylacetic acids. Maximal activity was found with hexanoic acid. High activities were obtained with benzoic acid having methyl, pentyl, and methoxy groups in the para- or meta-positions of the benzene ring. However, the enzyme was less active with valproic acid and ketoprofen. Salicylic acid exhibited no activity. The medium chain acyl-CoA synthetases from mouse and bovine liver mitochondria were subjected to in-gel tryptic digestion, followed by LC-MS/MS sequence analysis. The amino acid sequence of each tryptic peptide of mouse liver mitochondrial medium chain acyl-CoA synthetase differed from that from bovine liver mitochondria only in one or two amino acids. LC-MS/MS analysis provided the information about these differences in amino acid sequences. In addition, we compared the properties of this protein with the homologues from rat and bovine.
authors.affiliation: Faculty of Pharmaceutical Sciences, Kobe-Gakuin University, Ikawadani, Nishiku, Japan. kasuya@pharm.kobegakuin.ac.jp
authors.forename: Fumiyo
authors.initials: F
authors.lastname: Kasuya
authors.forename: Takao
authors.initials: T
authors.lastname: Tatsuki
authors.forename: Masayuki
authors.initials: M
authors.lastname: Ohta
authors.forename: Yuich
authors.initials: Y
authors.lastname: Kawai
authors.forename: Kazuo
authors.initials: K
authors.lastname: Igarashi
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002384:Catalysis; D002417:Cattle; D003066:Coenzyme A Ligases / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification* / Q000378:metabolism; D005227:Fatty Acids / Q000096:biosynthesis; D051379:Mice; D008930:Mitochondria, Liver / Q000201:enzymology* / Q000235:genetics; D024101:Mitochondrial Proteins; D008969:Molecular Sequence Data; D051381:Rats; D020539:Sequence Analysis, Protein; D017386:Sequence Homology, Amino Acid; D013045:Species Specificity; D013379:Substrate Specificity
chemical_list: D005227:Fatty Acids; D024101:Mitochondrial Proteins; C499057:Acsm1 protein, mouse; D003066:Coenzyme A Ligases