Cloning, expression and tissue distribution of a tetrameric form of pig carbonyl reductase.
In this study, we isolated a cDNA for tetrameric carbonyl reductase (CR) from pig heart. The pig CR showed high amino acid sequence identity (81%) with rabbit NADP(+)-dependent retinol dehydrogenase (NDRD). The purified recombinant pig CR and NDRD were about 100-kDa homotetramers and exhibited high reductase activity towards alkyl phenyl ketones, alpha-dicarbonyl compounds and all-trans-retinal. The identity of NDRD with the tetrameric CR was verified by protein sequencing of CR purified from rabbit heart. Both tetrameric CR and its mRNA were ubiquitously expressed in pig and rabbit tissues. The pig and rabbit enzymes belonged to the short-chain dehydrogenase/reductase family, and their sequences comprise a C-terminal SRL tripeptide, which is a variant of the type 1 peroxisomal targeting signal, SKL. Transfection of HeLa cells with vectors expressing pig CR demonstrated that the enzyme is localized in the peroxisomes. Thus, the tetrameric form of CR represents the first mammalian peroxisomal enzyme that reduces all-trans-retinal as the endogenous substrate.
Usami
Noriyuki
N
Biochemistry Laboratory, Gifu Pharmaceutical University, Mitahora-higashi, 502-8585, Gifu, Japan.
Ishikura
Shuhei
S
Abe
Hiroko
H
Nagano
Makoto
M
Uebuchi
Miki
M
Kuniyasu
Akihiko
A
Otagiri
Masaki
M
Nakayama
Hitoshi
H
Imamura
Yorishige
Y
Hara
Akira
A
D000429:Alcohol Oxidoreductases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D000449:Aldehyde Reductase; D000074408:Aldo-Keto Reductases; D000595:Amino Acid Sequence; D000818:Animals; D003001:Cloning, Molecular; D018076:DNA, Complementary; D008969:Molecular Sequence Data; D017386:Sequence Homology, Amino Acid; D013552:Swine; D014018:Tissue Distribution
D018076:DNA, Complementary; D000429:Alcohol Oxidoreductases; D000074408:Aldo-Keto Reductases; D000449:Aldehyde Reductase