Dephosphorylation of tubulin-bound guanosine triphosphate during microtubule assembly

1. Tubulin purified from porcine brain in the presence of GTP contained 0.16 mole of GDP and 0.73 mole of GTP per 60,000 g of protein. 2. Microtubules reconstituted from the purified tubulin contained 0.43 mole of GDP and 0.41 mole of GTP per 60,000 g of protein. Guanine nucleotide bound to the exchangeable site of tubulin was converted to GDP during microtubule assembly, while GTP at the non-exchangeable site remained intact. 3. Guanine nucleotide which had been bound to the exchangeable site of tubulin before microtubule assembly was also exchangeable during disassembly.