Cloning and expression of koala ('Phascolarctos cinereus') liver cytochrome P450 reductase

Title
Cloning and expression of koala ('Phascolarctos cinereus') liver cytochrome P450 reductase
Publication Date
2009
Author(s)
Kong, Sandra
Ngo, Suong NT
McKinnon, Ross A
Stupans, Ieva
Type of document
Journal Article
Language
en
Entity Type
Publication
Publisher
Elsevier Inc
Place of publication
United States of America
DOI
10.1016/j.cbpc.2009.01.010
UNE publication id
une:9142
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.
Link
Citation
Comparative Biochemistry and Physiology Part C: Toxicology & Pharmacology, 150(1), p. 1-9
ISSN
1878-1659
1532-0456
Start page
1
End page
9

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