Title: Radioactive assay for serine transhydroxymethylase
Abstract: A high-performance liquid chromatography (HPLC)-based fluorometric method for measuring serine hydroxymethyltransferase (SHMT) activity toward formation of serine and (6S)-H4PteGlun has been developed. In this method, serine formed by SHMT activity is reacted with 4-fluoro-7-nitro-2,1,3-benzoxadiazole (NBD-F) to form the fluorescent adduct NBD–serine. The fluorescent assay components are then separated by reversed-phase chromatography, and NBD–serine is quantified by comparison with standards. This method was used to determine the Km and kcat values for 5,10-CH2–H4PteGlu5 of an SHMT from Arabidopsis thaliana. These data represent the first determination of kinetic parameters for (6S)-5,10-CH2–H4PteGlu5 for an SHMT from any organism.
Publication Year: 1965
Publication Date: 1965-10-01
Language: en
Type: article
Indexed In: ['crossref']
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Cited By Count: 208
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