Deprotonations in the Reaction of Flavin-Dependent Thymidylate Synthase

Overview

Journal Biochemistry

Specialty Biochemistry

Date 2016 May 24

PMID 27214228

Citations 9

Authors

Frederick W Stull

Steffen M Bernard

Aparna Sapra

Janet L Smith

Erik R P Zuiderweg

Bruce A Palfey

Affiliations

Soon will be listed here.

Abstract

Many microorganisms use flavin-dependent thymidylate synthase (FDTS) to synthesize the essential nucleotide 2'-deoxythymidine 5'-monophosphate (dTMP) from 2'-deoxyuridine 5'-monophosphate (dUMP), 5,10-methylenetetrahydrofolate (CH2THF), and NADPH. FDTSs have a structure that is unrelated to the thymidylate synthase used by humans and a very different mechanism. Here we report nuclear magnetic resonance evidence that FDTS ionizes N3 of dUMP using an active-site arginine. The ionized form of dUMP is largely responsible for the changes in the flavin absorbance spectrum of FDTS upon dUMP binding. dUMP analogues also suggest that the phosphate of dUMP acts as the base that removes the proton from C5 of the dUMP-methylene intermediate in the FDTS-catalyzed reaction. These findings establish additional differences between the mechanisms of FDTS and human thymidylate synthase.

Citing Articles

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An enzymatic activation of formaldehyde for nucleotide methylation.

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