cDNA cloning and genomic organization of peroxisome proliferator-inducible long-chain acyl-CoA hydrolase from rat liver cytosol.
The cDNA for a peroxisome proliferator-inducible long-chain acyl-CoA hydrolase from rat liver cytosol, referred to as rLACH2, was isolated and its genomic structure was determined. The cDNA encoded a 419-amino-acid polypeptide with a calculated molecular weight of 46,011. Sequence analysis identified an active-site serine motif (Gly-x-Ser-x-Gly) common to carboxylesterases and lipases. When expressed in Escherichia coli, the cDNA directed expression of a protein immunoreactive to an anti-rLACH2 antibody with a molecular mass of 47 kDa, identical to that of purified rLACH2. Northern blot analysis showed marked induction of rLACH2 mRNA in the liver after feeding rats with di(2-ethylhexyl)phthalate, a peroxisome proliferator. The rLACH2 gene spanned about 19 kb and comprised 3 exons, the intron/exon boundaries of which were consistent with the donor/acceptor splice rule. A putative peroxisome proliferator response element (AGGTCATGGTTCA) was identified in the 5'-flanking region, suggesting the involvement of peroxisome proliferator-activated receptors in the regulation of rLACH2 gene expression.
Department of Clinical Biochemistry, Tokyo University of Pharmacy and Life Science, Japan. junymd@ps.toyaku.ac.jp
J
J
Yamada
K
K
Suga
T
T
Furihata
M
M
Kitahara
T
T
Watanabe
M
M
Hosokawa
T
T
Satoh
T
T
Suga
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D003001:Cloning, Molecular; D003600:Cytosol / Q000201:enzymology; D018076:DNA, Complementary; D004051:Diethylhexyl Phthalate / Q000494:pharmacology; D004790:Enzyme Induction; D004926:Escherichia coli; D005091:Exons; D015698:Genomic Library; D007438:Introns; D008099:Liver / Q000187:drug effects / Q000201:enzymology*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010170:Palmitoyl-CoA Hydrolase / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics*; D051381:Rats; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry; D012694:Serine
D018076:DNA, Complementary; D011994:Recombinant Proteins; D012694:Serine; D004051:Diethylhexyl Phthalate; D010170:Palmitoyl-CoA Hydrolase