采用荧光高效液相色谱法测定精氨酰转移酶活性
Assay of Arginyltransferase Activity by a Fluorescent HPLC Method.
作者信息
Takao Koichi
机构信息
Faculty of Pharmacy and Pharmaceutical Sciences, Josai University, Sakado, Saitama, Japan.
出版信息
Methods Mol Biol. 2023;2620:129-137. doi: 10.1007/978-1-0716-2942-0_18.
Syntheses of fluorescent substrate and product for arginyltransferase, N-aspartyl-4-dansylamidobutylamine (Asp4DNS) and N-arginylaspartyl-4-dansylamidobutylamine (ArgAsp4DNS), respectively, including their precursor 4-dansylamidobutylamine (4DNS), are described. Then, HPLC conditions are summarized for a baseline separation of the three compounds in 10 min. The present method, which permits the simultaneous determination of Asp4DNS, 4DNS, and ArgAsp4DNS (in eluting order), is advantageous in measuring arginyltransferase activity and detecting the unfavorable enzyme(s) in 105,000 × g supernatant of tissues to ensure accurate determination.
分别描述了用于精氨酰转移酶的荧光底物和产物N-天冬氨酰-4-丹磺酰胺基丁胺（Asp4DNS）和N-精氨酰天冬氨酰-4-丹磺酰胺基丁胺（ArgAsp4DNS）的合成方法，包括它们的前体4-丹磺酰胺基丁胺（4DNS）。然后总结了在10分钟内对这三种化合物进行基线分离的高效液相色谱条件。本方法能够同时测定Asp4DNS、4DNS和ArgAsp4DNS（按洗脱顺序），在测量组织105,000×g上清液中的精氨酰转移酶活性和检测不利酶方面具有优势，以确保准确测定。
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