A continuous spectrophotometric assay for Escherichia coli alanyl-transfer RNA synthetase

✍ Scribed by Siddhartha Roy

• Publisher: Elsevier Science
• Year: 1983
• Tongue: English
• Weight: 283 KB
• Volume: 133
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(83)90086-6

✦ Synopsis

A new continuous spectrophotometric assay is demonstrated for Escherichia coli alanyl-tRNA synthetase. It involves beta-gamma adenylyl imidophosphate as a substitute for ATP in the pyrophosphate exchange reaction. The net conversion of beta-gamma adenylyl imidophosphate to ATP can be linked to NADP reduction by hexokinase and glucose-6-P dehydrogenase catalyzed reactions, which can be monitored at 340 nm. This assay can be extended to other aminoacyl-tRNA synthetases which can use beta-gamma nonhydrolyzable analogs of ATP as an ATP substitute.