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Title: Cloning and expression of koala ('Phascolarctos cinereus') liver cytochrome P450 reductase
Contributor(s): Kong, Sandra (author); Ngo, Suong NT (author); McKinnon, Ross A (author); Stupans, Ieva  (author)
Publication Date: 2009
DOI: 10.1016/j.cbpc.2009.01.010
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Abstract: The cloning, expression and characterization of hepatic NADPH-cytochrome P450 reductase (CPR) from koala ('Phascolarctos cinereus') is described. Two 2059 bp koala liver CPR cDNAs, designated CPR1 and CPR2, were cloned by reverse transcription-polymerase chain reaction and rapid amplification of cDNA ends. The koala CPR cDNAs encode proteins of 678 amino acids and share 85% amino acid sequence identity to human CPR. Transfection of the koala CPR cDNAs into Cos-7 cells resulted in the expression of proteins, which were recognized by a goat-antihuman CPR antibody. The koala CPR1 and 2 cDNA-expressed enzymes catalysed cytochrome c reductase at the rates of 4.9 ± 0.5 and 2.6 ± 0.4 nmol/min/mg protein (mean ± SD, n = 3), respectively which were comparable to that of rat CPR cDNA-expressed enzyme. The apparent Km value for CPR activity in koala liver microsomes was 11.61 ± 6.01 µM, which is consistent with that reported for rat CPR enzyme. Northern analysis detected a CPR mRNA band of approximately 2.6 kb. Southern analysis suggested a single PCR gene across species. The present study provides primary molecular data regarding koala CPR1 and CPR2 genes in this unique marsupial species.
Publication Type: Journal Article
Source of Publication: Comparative Biochemistry and Physiology Part C: Toxicology & Pharmacology, 150(1), p. 1-9
Publisher: Elsevier Inc
Place of Publication: United States of America
ISSN: 1878-1659
Field of Research (FOR): 119999 Medical and Health Sciences not elsewhere classified
Socio-Economic Outcome Codes: 929999 Health not elsewhere classified
Peer Reviewed: Yes
HERDC Category Description: C1 Refereed Article in a Scholarly Journal
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