Characterization of indolamine N-acetyltransferase activity from the head ganglia of the American cockroach, Periplaneta americana
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概要
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More than 90% of indolamine N-acetyltransferase (NAT) activity was recovered in the supernatant of centrifuged homogenate of the head ganglia of the American cockroach, Periplaneta americana. NAT activity increased at temperatures of up to 40℃ with serotonin (5HT) and up to 45℃ with tryptamine (TP) as the substrates. The pH-activity curve with 5HT resembled that with TP, both showing two activity maxima, one at pH 7.0 and the other at 8.5-9.0. Na^+ and K^+ ions stimulated the enzymatic activity up to 200 mM, after which a further increase in the concentrations reduced the enzyme activity. Cu^<2+> suppressed the activity at least up to 0.5 mM. One millimolar Mg^<2+> stimulated the activity but a further increase in the concentration suppressed the activity. Ca^<2+> also stimulated the activity up to 2 mM, but a further increase suppressed the activity. The secondary plot of the ordinate intercepts of a series of Lineweaver-Burk plots against fixed concentrations of acetyl-CoA showed that the enzymatic mechanism of indolamine NAT from the head ganglia of P. americana proceeds according to the kinetics known as the ping pong Bi-Bi mechanism. The apparent K_ms and V_<max> with TP and acetyl-CoA were 19.3 μM and V_<max> 3.64 nmol product/mg protein/min, respectively, while the K_m for acetyl-CoA was 28.0 μM and V_<max> the same as that for TP. The K_m for 5HT was 106 μM, V_<max> being 2.16 nmol product/mg protein/min, while the K_m for acetyl-CoA was 19.7 μM, V_<max> being the same as that for 5HT.
- 1998-02-25
著者
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Takeda Makio
Laboratory Of Insect Science Faculty Of Agriculture Kobe University
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Sakamoto Toshiro
Fujisawa Pharmaceutical Ltd.
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Takeda Makio
Laboratory Of Entomology Faculty Of Agrieulture Kobe University
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Ichihara Naoyuki
Laboratory of Insect Science, Faculty of Agriculture, Kobe University
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Ichihara N
Kobe Univ. Kobe Jpn
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Ichihara Naoyuki
Laboratoire De Mathematiques Universite De Bretagne Occidentale
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