An HPLC-based fluorometric assay for serine hydroxymethyltransferase

We have developed a novel HPLC-based fluorometric assay for serine hydroxymethyltransferase activity. In this assay, the 5,10-CH2-H4PteGlu formed by serine hydroxymethyltransferase activity is reduced to 5-CH3-H4PteGlu using NaBH4. Then the fluorescent assay components are separated by reversed-phas...

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Bibliographic Details
Published inAnalytical biochemistry Vol. 375; no. 2; pp. 367 - 369
Main Authors Zhang, Yi, Sun, Kehan, Roje, Sanja
Format Journal Article
LanguageEnglish
Published United States Elsevier Inc 15.04.2008
Subjects
Arabidopsis - enzymology
Chromatography, High Pressure Liquid
Glycine Hydroxymethyltransferase - metabolism
Kinetics
Plant Leaves - enzymology
Spectrometry, Fluorescence
Tetrahydrofolates - metabolism
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Summary:We have developed a novel HPLC-based fluorometric assay for serine hydroxymethyltransferase activity. In this assay, the 5,10-CH2-H4PteGlu formed by serine hydroxymethyltransferase activity is reduced to 5-CH3-H4PteGlu using NaBH4. Then the fluorescent assay components are separated by reversed-phase chromatography under isocratic conditions and 5-CH3-H4PteGlu is quantified by comparison with standards. We show that this assay can be used to measure serine hydroxymethyltransferase activity at 10−8 to 10−3M (6R,S)-H4PteGlu.
Bibliography:ObjectType-Article-1
SourceType-Scholarly Journals-1
ObjectType-Feature-2
content type line 23
ISSN:0003-2697
1096-0309
DOI:10.1016/j.ab.2007.12.015