A convenient method for enzymatic synthesis of radiolabelled glucose-1,6-bisphosphate

✍ Scribed by Tom Hamborg Nielsen; Bente Wischmann; Birger Lindberg Møller

• Publisher: John Wiley and Sons
• Year: 1995
• Tongue: French
• Weight: 309 KB
• Volume: 36
• Category: Article
• ISSN: 0022-2135
• DOI: 10.1002/jlcr.2580360710

✦ Synopsis

Abstract

A convenient method to synthesize radiolabelled glucose‐1,6‐bisphosphate (Glc‐1,6‐P~2~) of high specific activity and in a high yield is reported. The method is based on enzymatic formation of glucose‐6‐P from glucose and ATP using hexokinase. The labelled glucose‐6‐P is then converted to Glc‐1,6‐P~2~ through an equilibrium reaction catalyzed by phosphoglucomutase. The two enzymatic steps and TLC separations necessary to afford radiochemically pure Glc‐1,6‐P~2~ can be completed in one day. The outlined experimental procedure details the synthesis of [^33^P]Glc‐1,6‐P~2~ evenly labelled at the carbon 1 and the carbon 6 position. However the commercial availability of γ‐^33^P and γ‐^32^P labelled ATP and of glucose labelled with ^3^H, ^14^C and ^13^C provides the option to label glucose‐1,6‐bisphosphate with any of these isotopes.