Purification, molecular cloning, and genomic organization of human brain long-chain acyl-CoA hydrolase.
An acyl-CoA hydrolase, referred to as hBACH, was purified from human brain cytosol. The enzyme had a molecular mass of 100 kDa and 43-kDa subunits, and was highly active with long-chain acyl-CoAs, e.g. a maximal velocity of 295 micromol/min/mg and K(m) of 6.4 microM for palmitoyl-CoA. Acyl-CoAs with carbon chain lengths of C(8-18) were also good substrates. In human brain cytosol, 85% of palmitoyl-CoA hydrolase activity was titrated by an anti-BACH antibody, which accounted for over 75% of the enzyme activity found in the brain tissue. The cDNA isolated for hBACH, when expressed in Escherichia coli, directed the expression of palmitoyl-CoA hydrolase activity and a 44-kDa protein immunoreactive to the anti-BACH antibody, which in turn neutralized the hydrolase activity. The hBACH cDNA encoded a 338-amino acid sequence which was 95% identical to that of a rat homolog. The hBACH gene spanned about 130 kb and comprised 9 exons, and was mapped to 1p36.2 on the cytogenetic ideogram. These findings indicate that the long-chain acyl-CoA hydrolase present in the brain is well conserved between man and the rat, suggesting a conserved role for this enzyme in the mammalian brain, and enabling genetic studies on the functional analysis of acyl-CoA hydrolase.
Yamada
J
J
Department of Clinical Biochemistry, Tokyo University of Pharmacy and Life Science, Hachioji, Tokyo, 192-0392, Japan. junymd@ps.toyaku. ac.jp
Kurata
A
A
Hirata
M
M
Taniguchi
T
T
Takama
H
H
Furihata
T
T
Shiratori
K
K
Iida
N
N
Takagi-Sakuma
M
M
Watanabe
T
T
Kurosaki
K
K
Endo
T
T
Suga
T
T
D000328:Adult; D000368:Aged; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001921:Brain / Q000201:enzymology*; D003001:Cloning, Molecular; D003600:Cytosol / Q000201:enzymology; D006801:Humans; D008297:Male; D008875:Middle Aged; D008969:Molecular Sequence Data; D010170:Palmitoyl-CoA Hydrolase / Q000235:genetics / Q000378:metabolism*; D051381:Rats
D010170:Palmitoyl-CoA Hydrolase