A modified enzymatic-isotopic microassay for serotonin (5HT) using 5HT-N-acetyltransferase partially purified from Drosophila.
A modified enzymatic-isotopic microassay for serotonin (5HT) using 5HT-N-acetyltransferase partially purified from Drosophila.
复制标题
使用从果蝇中部分纯化的 5HT-N-乙酰转移酶对血清素 (5HT) 进行改进的酶同位素微量测定。
DOI：
10.1016/0024-3205(83)90676-8
复制
发表时间：
1983
期刊：
影响因子：
6.1
通讯作者：
Jimenez,A
中科院分区：
文献类型：
--
作者：
Walker,RF;Friedman,DW;Jimenez,A
摘要血清素-N-乙酰转移酶（EC 2.3。1.5），部分纯化的黑腹果蝇（DNAT）取代大鼠肝酶（RNAT）在以前发表的5-羟色胺（5-HT）的放射酶法测定。目的是确定该修改是否会增加原始测定的灵敏度和可靠性。与RNAT相比，DNAT的活性高3-4倍;它缺乏二级5 HTP脱羧酶活性;并且它更稳定。在改进的测定条件下，下丘脑组织提取物形成的唯一放射性产物是3 H-褪黑激素;其可以从低至15 pg的5-羟色胺底物中测量。因此，用DNAT代替RNAT改进了原来的放射酶测定法，允许测量来自个体动物的下丘脑核中的内源性5-HT。
Abstract Serotonin-N-acetyltransferase (EC 2.3. 1.5), partially purified from Drosophila melanogaster (DNAT) was substituted for rat liver enzyme (RNAT) in a previously published radioenzymatic assay for serotonin (5HT). The purpose was to determine if this modification would increase the sensitivity and reliability of the original assay. Compared with RNAT, DNAT is 3–4 times more active; it lacks secondary 5HTP decarboxylase activity; and it is more stable. Under conditions of the modified assay, the only radioactive product formed from hypothalamic tissue extracts is 3 H-melatonin; which can be measured from as little as 15 pg of serotonin substrate. Thus, substitution of DNAT for RNAT improves the original radioenzymatic assay allowing measurement of endogenous 5HT in hypothalamic nuclei from individual animals.