Cloning and characterization of insect arylalkylamine N-acetyltransferase from Bombyx mori.
Arylalkylamine N-acetyltransferase (AANAT) catalyzes N-acetylation of arylarkylamines. A cDNA of Bombyx mori insect AANAT (Bm-iAANAT) was found by searching an expressed-sequence tag (EST) database of B. mori (SilkBase). The cDNA encoded a 261 amino acid protein. The mRNA of Bm-iAANAT was expressed in eggs, larvae, adults and various tissues. Recombinant Bm-iAANAT protein was expressed in Sf9 cells by a baculovirus expression system. The AANAT activity of Bm-iAANAT was inhibited by high concentrations (over 0.01 mM) of tryptamine used as a substrate. The Bm-iAANAT acetylated tryptamine, serotonin, dopamine, octopamine, tyramine and norepinephrine. This is the first report of a cloned AANAT that acetylated norepinephrine. These results suggest that Bm-iAANAT is a novel member of insect AANAT family with unique kinetic properties and a broad substrate range.
Tsugehara
Taketo
T
Division of Molecular Science, Graduate School of Science and Technology, Kobe University, Kobe, Hyogo, Japan.
Iwai
Sachio
S
Fujiwara
Yoshihiro
Y
Mita
Kazuei
K
Takeda
Makio
M
D000818:Animals; D047068:Arylalkylamine N-Acetyltransferase / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics*; D016367:Baculoviridae; D001483:Base Sequence; D012831:Bombyx / Q000201:enzymology* / Q000235:genetics*; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D015870:Gene Expression; D019476:Insect Proteins / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics*; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D013379:Substrate Specificity
D018076:DNA, Complementary; D019476:Insect Proteins; D011994:Recombinant Proteins; D047068:Arylalkylamine N-Acetyltransferase