一种通用板格式，用于提高监测多种氨酰基转移RNA合成酶活性的检测通量。
A universal plate format for increased throughput of assays that monitor multiple aminoacyl transfer RNA synthetase activities.
作者信息
Beebe Kirk, Waas William, Druzina Zhanna, Guo Min, Schimmel Paul
机构信息
Department of Molecular Biology and Chemistry and Skaggs Institute for Chemical Biology, Scripps Research Institute, La Jolla, CA 92037, USA.
出版信息
Anal Biochem. 2007 Sep 1;368(1):111-21. doi: 10.1016/j.ab.2007.05.013. Epub 2007 May 18.
Abstract
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.
摘要
氨酰转移核糖核酸（tRNA）合成酶是备受深入研究的酶类，因为它们在遗传密码的确立中具有重要性，且与疾病和医学相关。在该领域的发展过程中，人们开发了多种检测方法。尽管有许多创新，但放射性检测方法（最早开发的检测方法之一）的灵敏度、简便性和可靠性确保了它们仍在持续使用。这些检测方法可测量四种活性：活性位点滴定、氨基酸活化、氨酰化和转移后编辑（去酰化）。为了在保持这些检测方法优势的同时提高通量、减少浪费并改善数据质量，人们开发了一种通用的96孔滤板形式。这种形式便于对广泛研究的所有四种活性进行检测。
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