|
type |
Journal Article |
authors |
Kruschwitz, H.; Ren, S.; Di Salvo, M.; Schirch, V. |
title |
Expression, purification, and characterization of human cytosolic serine hydroxymethyltransferase |
journal |
Protein Expr Purif |
Activity |
2.1.2.1 |
Family |
2.1.2.1 |
sel |
selected |
ui |
8527925 |
year |
(1995) |
volume |
6 |
number |
4 |
pages |
411-6 |
| |
keywords |
Amino Acid Sequence |
abstract |
A cDNA which codes for human cytosolic serine hydroxymethyltransferase (Garrow et al., 1993, J. Biol. Chem. 268, 11910-11916) has been cloned into a pT7-7 vector as a NdeI-EcoRI insert. HMS174 (de3) cells were transformed with this plasmid and, after induction with isopropyl thiogalactoside, expressed a catalytically active serine hydroxymethyltransferase. The enzyme was purified and shown to be the expressed human enzyme by N-terminal amino acid sequencing. About 225 mg of pure enzyme can be obtained from a 20-liter culture. Spectral characteristics of the bound pyridoxal phosphate were essentially identical to the spectral properties of rabbit cytosolic serine hydroxymethyltransferase. Kinetic constants for the natural substrates L-serine and tetrahydrofolate were also similar to the values obtained previously for the rabbit cytosolic enzyme. |
last changed |
2009/01/12 12:47 |
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