An HPLC-based fluorometric assay for serine hydroxymethyltransferase

Yi Zhang; Kehan Sun; Sanja Roje

doi:10.1016/j.ab.2007.12.015

An HPLC-based fluorometric assay for serine hydroxymethyltransferase

Anal Biochem. 2008 Apr 15;375(2):367-9. doi: 10.1016/j.ab.2007.12.015. Epub 2007 Dec 23.

Authors

Yi Zhang¹, Kehan Sun, Sanja Roje

Affiliation

¹ Institute of Biological Chemistry, Washington State University, Pullman, WA 99164, USA.

PMID: 18194659
DOI: 10.1016/j.ab.2007.12.015

Abstract

We have developed a novel HPLC-based fluorometric assay for serine hydroxymethyltransferase activity. In this assay, the 5,10-CH(2)-H(4)PteGlu formed by serine hydroxymethyltransferase activity is reduced to 5-CH(3)-H(4)PteGlu using NaBH(4). Then the fluorescent assay components are separated by reversed-phase chromatography under isocratic conditions and 5-CH(3)-H(4)PteGlu is quantified by comparison with standards. We show that this assay can be used to measure serine hydroxymethyltransferase activity at 10(-8) to 10(-3)M (6R,S)-H(4)PteGlu.

Publication types

Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

Arabidopsis / enzymology
Chromatography, High Pressure Liquid
Glycine Hydroxymethyltransferase / metabolism*
Kinetics
Plant Leaves / enzymology
Spectrometry, Fluorescence
Tetrahydrofolates / metabolism

Substances

Tetrahydrofolates
5,10-methylenetetrahydrofolic acid
Glycine Hydroxymethyltransferase