A universal plate format for increased throughput of assays that monitor multiple aminoacyl transfer RNA synthetase activities

被引:35
作者
Beebe, Kirk
Waas, William
Druzina, Zhanna
Guo, Min
Schimmel, Paul [1 ]
机构
[1] Scripps Res Inst, Dept Mol Biol & Chem, La Jolla, CA 92037 USA
[2] Scripps Res Inst, Skaggs Inst Chem Biol, La Jolla, CA 92037 USA
关键词
transfer RNA; filter plate; editing; scintillation counting; aminoacyl tRNA synthetase; ATP-PPi exchange; aminoacylation; charging;
D O I
10.1016/j.ab.2007.05.013
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Aminoacyl transfer RNA (tRNA) synthetases are intensely studied enzymes because of their importance in the establishment of the genetic code and their connection to disease and medicine. During the advancement of this field, several assays were developed. Despite many innovations, the sensitivity, simplicity, and reliability of the radiometric assays (which were among the first to be developed) have ensured their continued use. Four activities are measured by these assays: active site titration, amino acid activation, aminoacylation, and posttransfer editing (deacylation). In an effort to maintain the advantage of these assays while enhancing throughput, reducing waste, and improving data quality, a universal 96-well filter plate format was developed. This format facilitates the assays for all four of the widely studied activities. (c) 2007 Elsevier Inc. All rights reserved.
引用
收藏
页码:111 / 121
页数:11
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