title: An HPLC-based fluorometric assay for serine hydroxymethyltransferase.
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.
authors.affiliation: Institute of Biological Chemistry, Washington State University, Pullman, WA 99164, USA.
authors.forename: Yi
authors.initials: Y
authors.lastname: Zhang
authors.forename: Kehan
authors.initials: K
authors.lastname: Sun
authors.forename: Sanja
authors.initials: S
authors.lastname: Roje
mesh_terms: 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
chemical_list: D013763:Tetrahydrofolates; C013123:5,10-methylenetetrahydrofolic acid; D012696:Glycine Hydroxymethyltransferase