The stereochemical course of thiophosphoryl group transfer catalyzed by T4 polynucleotide kinase.
The stereochemical course of the phosphoryl transfer reaction catalyzed by T4 polynucleotide kinase has been determined using the chiral ATP analog, (Sp)-adenosine-5'-(3-thio-3-[18O]triphosphate). T4 polynucleotide kinase catalyzes the transfer of the gamma-thiophosphoryl group of (Sp)-adenosine-5'-(3-thio-3-[18O]triphosphate) to the 5'-hydroxyl group of ApA to give the thiophosphorylated dinucleotide adenyl-5'-[18O]phosphorothioate-(3'-5')adenosine. A sample of adenyl-5'-[18O]phosphorothioate-(3'-5')adenosine was subjected to venom phosphodiesterase digestion. The resulting adenosine-5'-[18O]phosphorothioate was shown to have the Rp configuration, thus indicating that the thiophosphoryl transfer reaction occurs with overall inversion of configuration of phosphorus.
D000227:Adenine Nucleotides / Q000378:metabolism; D000249:Adenosine Monophosphate / Q000031:analogs & derivatives / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000031:analogs & derivatives* / Q000378:metabolism; D002384:Catalysis; D015226:Dinucleoside Phosphates; D008401:Gas Chromatography-Mass Spectrometry; D007527:Isoenzymes; D008968:Molecular Conformation; D043264:Phosphodiesterase I; D010727:Phosphoric Diester Hydrolases; D010770:Phosphotransferases / Q000378:metabolism*; D011116:Polynucleotide 5'-Hydroxyl-Kinase / Q000378:metabolism*; D013604:T-Phages / Q000201:enzymology*; D013873:Thionucleotides / Q000378:metabolism*
D000227:Adenine Nucleotides; D015226:Dinucleoside Phosphates; D007527:Isoenzymes; D013873:Thionucleotides; C006412:adenylyl-(3'-5')-adenosine; C022571:adenosine 5'-O-(3-thiotriphosphate); D000249:Adenosine Monophosphate; C030359:adenyl-5'-O-phosphorothioate-(3'-5')adenosine; D000255:Adenosine Triphosphate; D010770:Phosphotransferases; D011116:Polynucleotide 5'-Hydroxyl-Kinase; D010727:Phosphoric Diester Hydrolases; D043264:Phosphodiesterase I