Transition State Chirality and Role of the Vicinal Hydroxyl in the Ribosomal Peptidyl Transferase Reaction

Overview

Journal Biochemistry

Specialty Biochemistry

Date 2008 Aug 5

PMID 18672893

Citations 3

Authors

Kevin S Huang

Nicolas Carrasco

Emmanuel Pfund

Scott A Strobel

Affiliations

Soon will be listed here.

Abstract

The ribosomal peptidyl transferase is a biologically essential catalyst responsible for protein synthesis. The reaction is expected to proceed through a transition state approaching tetrahedral geometry with a specific chirality. To establish that stereospecificity, we synthesized two diastereomers of a transition state inhibitor with mimics for each of the four ligands around the reactive chiral center. Preferential binding of the inhibitor that mimics a transition state with S chirality establishes the spatial position of the nascent peptide and the oxyanion and places the amine near the critical A76 2'-OH group on the P-site tRNA. Another inhibitor series with 2'-NH 2 and 2'-SH substitutions at the critical 2'-OH group was used to test the neutrality of the 2'-OH group as predicted if the hydroxyl functions as a proton shuttle in the transition state. The lack of significant pH-dependent binding by these inhibitors argues that the 2'-OH group remains neutral in the transition state. Both of these observations are consistent with a proton shuttle mechanism for the peptidyl transferase reaction.

Citing Articles

Different substrate-dependent transition states in the active site of the ribosome.

Kuhlenkoetter S, Wintermeyer W, Rodnina M Nature. 2011; 476(7360):351-4.

PMID: 21804565 DOI: 10.1038/nature10247.

A two-step chemical mechanism for ribosome-catalysed peptide bond formation.

Hiller D, Singh V, Zhong M, Strobel S Nature. 2011; 476(7359):236-9.

PMID: 21765427 PMC: 3154986. DOI: 10.1038/nature10248.

A structural view on the mechanism of the ribosome-catalyzed peptide bond formation.

Simonovic M, Steitz T Biochim Biophys Acta. 2009; 1789(9-10):612-23.

PMID: 19595805 PMC: 2783306. DOI: 10.1016/j.bbagrm.2009.06.006.

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