A transition state analog for phosphate diester cleavage catalyzed by a small enzyme-like metal ion complex.

The values of K(i) for methylphosphate dianion (MP(2-)) inhibition of the cleavage of 2-hydroxypropyl-4-nitrophenyl phosphate (HpPNP) catalyzed by 1,3-bis(1,4,7-triazacyclonon-1-yl)-2-hydroxypropane (Zn(2)(1)(H(2)O)) approach a small limiting value of 6 microM at pH<pK(a)=7.8 for deprotonation of the catalyst to form Zn(2)(1)(HO(-)). There is a 1600-fold difference in the affinity of a phosphate diester monoanion (diethylphosphate) and phosphate monester dianion (MP(2-)) for Zn(2)(1)(H(2)O). This suggests that the latter is an analog for the transition state dianion for the cleavage reaction of HpPNP and other phosphate diesters. The observation that this transition state analog binds selectively to Zn(2)(1)(H(2)O) provides strong evidence that this is the active form of the catalyst which binds selectively to the ionized substrate. The efficiency of catalysis of the cleavage of phosphate diesters by Zn(2)(1)(H(2)O) and proteins is compared.