Journal article
A high-performance liquid chromatography-based fluorometric method for assaying serine hydroxymethyltransferase toward serine formation
Analytical biochemistry, Vol.409(1), pp.156-158
2011
Handle:
https://hdl.handle.net/2376/109348
PMID: 20946865
Abstract
A high-performance liquid chromatography (HPLC)-based fluorometric method for measuring serine hydroxymethyltransferase (SHMT) activity toward formation of serine and (6
S)-H
4PteGlu
n
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
K
m and
k
cat values for 5,10-CH
2–H
4PteGlu
5 of an SHMT from
Arabidopsis thaliana. These data represent the first determination of kinetic parameters for (
6S)-5,10-CH
2–H
4PteGlu
5 for an SHMT from any organism.
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Details
- Title
- A high-performance liquid chromatography-based fluorometric method for assaying serine hydroxymethyltransferase toward serine formation
- Creators
- Zhaoyang WeiSanja Roje
- Publication Details
- Analytical biochemistry, Vol.409(1), pp.156-158
- Academic Unit
- Biological Chemistry, Institute of
- Publisher
- Elsevier Inc
- Identifiers
- 99900547251501842
- Language
- English
- Resource Type
- Journal article