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Determination of kinetic properties of serotonin‐N‐acetyltransferase in bovine pineal gland using HPLC with fluorimetric detection
Author(s) -
Fajardo Natalia,
Abreu Pedro,
Alonso Rafael
Publication year - 1992
Publication title -
journal of pineal research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.881
H-Index - 131
eISSN - 1600-079X
pISSN - 0742-3098
DOI - 10.1111/j.1600-079x.1992.tb00058.x
Subject(s) - tryptamine , pineal gland , serotonin , chromatography , melatonin , enzyme , chemistry , high performance liquid chromatography , biochemistry , nat , biology , endocrinology , computer network , receptor , computer science
The determination of serotonin‐N‐acetyltransferase (NAT) activity in the bovine pineal gland and other rat tissues was based upon the separation and detection of N‐acetyltryptamine formed from tryptamine and acetyl CoA by means of high performance liquid chromatography with fluorimetric detection. In the bovine pineal the enzyme exhibited a Km value of 31.45 ± 4.98 μM and a Vmax value of 30.90 ± 1.18 pmol N‐acetyltryptamine/min/mg protein for tryptamine, and a Km value of 28.72 ± 7.50 μM and a Vmax value of 25.90 μ 1.50 pmol N‐acetyltryptamine/min/mg protein for acetyl CoA. The present method is simple, allows the determination of NAT activity from a variety of enzyme sources, has application to pharmacological studies of NAT regulation in tissue cultures, and provides an alternative to current radioenzymatic assays.