A Continuous Spectrophotometric Assay for the Aminoacylation of Transfer RNA by Alanyl-Transfer RNA Synthetase

✍ Scribed by M.X. Wu; K.A.W. Hill

• Publisher: Elsevier Science
• Year: 1993
• Tongue: English
• Weight: 272 KB
• Volume: 211
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1006/abio.1993.1276

✦ Synopsis

A continuous spectrophotometric assay is described for the aminoacylation reaction catalyzed by Escherichia coli alanyl-transfer RNA synthetase. The assay is based on coupling the alanyl-tRNA synthetase-dependent formation of AMP to the lactate dehydrogenase oxidation of NADH. Oxidation of NADH, as monitored at 340 nm, is shown to be stoichiometric with the formation of alanyl-tRNA(Ala). This assay will facilitate the rapid accumulation and analysis of kinetic data for alanyl-tRNA synthetase and should be applicable to aminoacyl-tRNA synthetases in general.