Please use this identifier to cite or link to this item: https://hdl.handle.net/2440/11268
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Type: Journal article
Title: Cloning, sequencing and expression of rat liver pyruvate carboxylase
Author: Jitrapakdee, S.
Booker, G.
Cassady, A.
Wallace, J.
Citation: Biochemical Journal, 1996; 316(2):613-637
Publisher: Portland Press
Issue Date: 1996
ISSN: 0264-6021
1470-8728
Statement of
Responsibility: 
Sarawut Jitrapakdee, Grant W. Booker, A. Ian Cassady and John C. Wallace
Abstract: Overlapping 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.
Keywords: Liver
Kidney
Animals
Rats
Chymotrypsin
Ligases
Pyruvate Carboxylase
Carbon-Nitrogen Ligases
DNA, Complementary
Cloning, Molecular
Polymerase Chain Reaction
Sequence Alignment
Gene Expression
Binding Sites
Amino Acid Sequence
Base Sequence
Consensus Sequence
Protein Conformation
Models, Molecular
Molecular Sequence Data
Rights: The Biochemical Society, London © 1996
DOI: 10.1042/bj3160631
Published version: http://www.biochemj.org/bj/316/bj3160631.htm
Appears in Collections:Aurora harvest 2
Biochemistry publications

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