Corroboration of Zn(ii)-Mg(ii)-tertiary structure interplays essential for the optimal catalysis of a phosphorothiolate thiolesterase ribozyme.

ABSTRACT: The TW17 ribozyme, a catalytic RNA selected from a pool of artificial RNA, is specific for the Zn²⁺-dependent hydrolysis of a phosphorothiolate thiolester bond. Here, we describe the organic synthesis of both guanosine α-thio-monophosphate and the substrates required for selecting and characterizing the TW17 ribozyme, and for deciphering the catalytic mechanism of the ribozyme. By successively substituting the substrate originally conjugated to the RNA pool with structurally modified substrates, we demonstrated that the TW17 ribozyme specifically catalyzes phosphorothiolate thiolester hydrolysis. Metal titration studies of TW17 ribozyme catalysis in the presence of Zn²⁺ alone, Zn²⁺ and Mg²⁺, and Zn²⁺ and [Co(NH₃)₆

SUBMITTER: Wang TP 

PROVIDER: S-EPMC9086351 | biostudies-literature | 2018 Sep

REPOSITORIES: biostudies-literature