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

Academic Article
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Overview

authors

  • Beebe, K.
  • Waas, W.
  • Druzina, Z.
  • Guo, Min
  • Schimmel, Paul

publication date

  • September 2007

journal

  • Analytical Biochemistry  Journal

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.

subject areas

  • Amino Acyl-tRNA Synthetases
  • Aminoacylation
  • Binding Sites
  • Models, Biological
  • Radiometry
  • Scintillation Counting
  • Sensitivity and Specificity
  • Time Factors
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Research

keywords

  • ATP-PPi exchange
  • aminoacyl tRNA synthetase
  • aminoacylation
  • charging
  • editing
  • filter plate
  • scintillation counting
  • transfer RNA
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Identity

PubMed Central ID

  • PMC3833075

International Standard Serial Number (ISSN)

  • 0003-2697

Digital Object Identifier (DOI)

  • 10.1016/j.ab.2007.05.013

PubMed ID

  • 17603003
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Additional Document Info

start page

  • 111

end page

  • 121

volume

  • 368

issue

  • 1

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