Microtubule assembly with the guanosine 5'-diphosphate analogue 2',3'-dideoxyguanosine 5'-diphosphate.
The GDP analogue 2',2'-dideoxyguanosine 5'-diphosphate (ddGDP) supports efficient tubulin polymerization. Microtubule-associated protein (MAP) dependent microtubule assembly occurs in 0.1 M 2-(N-morpholino)-ethanesulfonate, and sheets of parallel protofilaments are formed in 1.0 M glutamate without MAPs. The nucleotide is bound to tubulin in the course of polymerization, presumably in the exchangeable GTP site. The ddGDP is not hydrolyzed, however, and is completely stable in the reaction mixture. Nor was the nonexchangeable GTP bound to tubulin hydrolyzed in ddGDP-supported polymerization: equivalent amounts of GTP remained associated with polymerized tubulin after polymerization with either ddGDP or GTP. Higher concentrations of ddGDP than GTP were required under all conditions. Nevertheless, under optimum conditions for the ddGDP-supported reaction, polymerization began with a shorter lag period and both the rate and extent of polymerization were greater with ddGDP than with GTP. The MAP-dependent reaction with ddGDP is temperature dependent, cold reversible, and inhibited by calcium and antimitotic drugs. It differs from the GTP-supported reaction in being most vigorous at minimal Mg2+ concentrations and exquisitely sensitive to GDP inhibition.
D000818:Animals; D001921:Brain / Q000378:metabolism; D002417:Cattle; D003848:Deoxyguanine Nucleotides / Q000494:pharmacology*; D054306:Dideoxynucleotides; D004926:Escherichia coli / Q000378:metabolism; D007700:Kinetics; D008854:Microscopy, Electron; D008869:Microtubule-Associated Proteins; D008870:Microtubules / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure*; D020653:Peptide Elongation Factor G; D010445:Peptide Elongation Factors / Q000378:metabolism; D011506:Proteins / Q000378:metabolism; D012270:Ribosomes / Q000378:metabolism; D014404:Tubulin / Q000378:metabolism
D003848:Deoxyguanine Nucleotides; D054306:Dideoxynucleotides; D008869:Microtubule-Associated Proteins; D020653:Peptide Elongation Factor G; D010445:Peptide Elongation Factors; D011506:Proteins; D014404:Tubulin; C039033:2',3'-dideoxyguanosine 5'-diphosphate