title: The stereochemical course of thiophosphoryl group transfer catalyzed by T4 polynucleotide kinase.
abstract: 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.
mesh_terms: 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*
chemical_list: 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