The polymerization of 8-Azaguanosine 5′-diphosphate by polynucleotide phosphorylase

Add time:09/07/2019    Source:sciencedirect.com

8-Azaguanosine 5′-diphosphate serves as a substrate for polynucleotide phosphorylase. The terminal phosphate of the nucleoside diphosphate undergoes an exchange with [32P]orthophosphate in the presence of an enzyme preparation from Micrococcus lysodeikticus. This exchange is stimulated 5-fold by the addition of primer pApApA. In comparison, a similar 32P exchange with GDP is stimulated 25-fold in this system.In the presence of primer pApA and pApApA, 8-azaguanosine 5′-diphosphate is polymerized by highly purified polynucleotide phosphorylase isolated from Azotobacter agilis. An average chain-length of 10 mononucleotide units has been obtained.Exposure of poly azaguanylic acid to exhaustive digestion by pancreatic ribonuclease and takadiastase-T1-ribonuclease results in virtually complete degradation, the primary products being AzaG-2′,3′-P (cyclic), AzaG-3′(2′)-P (non-cyclic) and AzaGAzaG-2′,3′-P (cyclic).

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