title: Adipose pyruvate carboxylase: amino acid sequence and domain structure deduced from cDNA sequencing.
abstract: The complete amino acid sequence of 3T3-L1 adipocyte pyruvate carboxylase (PC) [pyruvate:carbon-dioxide ligase (ADP-forming), EC 6.4.1.1] has been deduced from sequencing overlapping cDNA clones obtained from an adipocyte cDNA library constructed in the lambda Zap vector. The encoding mRNA for PC promoter contains 4067 nt, including a 3534-nt coding sequence and noncoding regions of 100 and 433 nt at the 5' and 3' ends, respectively. The biotinylated lysine of the encoded PC promoter (1178 amino acids with a calculated M(r) of apocarboxylase = 129,784) is located 35 residues from the COOH-terminal end and, as in most other biotin enzymes, is in the consensus sequence AMKM. The adipocyte PC is closely similar (53% identity) to the yeast enzyme and contains different segments that are homologous with regions from the biotin carboxylase component of Escherichia coli acetyl-CoA carboxylase, the keto acid-binding subunits of Propionibacterium shermanii oxaloacetate transcarboxylase and Klebsiella pneumoniae oxaloacetate decarboxylase, and to the biotin carboxyl-carrier protein of the bacterial biotin enzymes. In addition to the putative mitochondrial targeting signal, functional domains are readily identifiable in the sequence and are in the following order: biotin carboxylase-carboxyltransferase-biotin carboxyl-carrier protein, as proposed for yeast PC.
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Miami School of Medicine, FL 33101.
authors.forename: J
authors.initials: J
authors.lastname: Zhang
authors.forename: W L
authors.initials: WL
authors.lastname: Xia
authors.forename: K
authors.initials: K
authors.lastname: Brew
authors.forename: F
authors.initials: F
authors.lastname: Ahmad
mesh_terms: D016475:3T3 Cells; D000273:Adipose Tissue / Q000201:enzymology*; D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics; D051379:Mice; D008969:Molecular Sequence Data; D011766:Pyruvate Carboxylase / Q000737:chemistry / Q000235:genetics* / Q000648:ultrastructure; D016415:Sequence Alignment
chemical_list: D004247:DNA; D011766:Pyruvate Carboxylase