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-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.
Zhang
Yi
Y
Institute of Biological Chemistry, Washington State University, Pullman, WA 99164, USA.
Sun
Kehan
K
Roje
Sanja
S
D017360:Arabidopsis / Q000201:enzymology; D002851:Chromatography, High Pressure Liquid; D012696:Glycine Hydroxymethyltransferase / Q000378:metabolism*; D007700:Kinetics; D018515:Plant Leaves / Q000201:enzymology; D013050:Spectrometry, Fluorescence; D013763:Tetrahydrofolates / Q000378:metabolism
D013763:Tetrahydrofolates; C013123:5,10-methylenetetrahydrofolic acid; D012696:Glycine Hydroxymethyltransferase