Jitrapakdee, S.Booker, G.Cassady, A.Wallace, J.2006-07-052006-07-051996Biochemical Journal, 1996; 316(2):613-6370264-60211470-8728http://hdl.handle.net/2440/11268Overlapping clones encoding rat liver pyruvate carboxylase (PC) have been isolated by screening a liver cDNA library and by performing rapid amplification of cDNA ends polymerase chain reaction on total liver RNA. The sequence of rat PC cDNA contains an open reading frame of 3537 nucleotides encoding a polypeptide of 1178 amino acids with a calculated Mr of 129848. This is flanked by a 5´ untranslated region of 66 bp and a 3´ untranslated region of 421 bp including the poly(A) tail. The inferred protein sequence is 96.6% identical with mouse and 96.3% identical with human PCs, 68.4% identical with mosquito PC and 53.5% identical with yeast PC isoenzymes PC1 and PC2. On the basis of partial proteolysis and sequence homology with PC from other organisms (yeast, mosquito, mouse and human) and with other biotin enzymes, three functional domains, namely the biotin carboxylation domain, the transcarboxylation domain and the biotinyl domain, have been identified. Comparison with the known structure of the biotin carboxylase subunit of Escherichia coli acetyl-CoA carboxylase [Waldrop, Rayment and Holden (1994) Biochemistry 33, 10249–10256] highlights the functional importance of 11 highly conserved residues. Northern analysis revealed that PC mRNA is highly expressed in rat liver, kidney, adipose tissue and brain, moderately expressed in heart, adrenal gland and lactating mammary gland, and expressed at a low level in spleen and skeletal muscle.enThe Biochemical Society, London © 1996LiverKidneyAnimalsRatsChymotrypsinLigasesPyruvate CarboxylaseCarbon-Nitrogen LigasesDNA, ComplementaryCloning, MolecularPolymerase Chain ReactionSequence AlignmentGene ExpressionBinding SitesAmino Acid SequenceBase SequenceConsensus SequenceProtein ConformationModels, MolecularMolecular Sequence DataJournal article0030004421001996540810.1042/bj3160631A1996UT232000382-s2.0-002994363168427Booker, G. [0000-0001-7207-4699]