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B6db references: 8527925

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
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

B6db references