EXPRESSION, PURIFICATION, AND CHARACTERIZATION OF HUMAN CYTOSOLIC SERINE HYDROXYMETHYLTRANSFERASE

被引：23

作者：

KRUSCHWITZ, H ^{[1
]}

REN, SL ^{[1
]}

DISALVO, M ^{[1
]}

SCHIRCH, V ^{[1
]}

机构：

[1] VIRGINIA COMMONWEALTH UNIV,DEPT BIOCHEM & MOLEC BIOPHYS,RICHMOND,VA 23298

来源：

PROTEIN EXPRESSION AND PURIFICATION | 1995年 / 6卷 / 04期

关键词：

D O I：

10.1006/prep.1995.1055

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

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. (C) 1995 Academic Press, Inc.

引用

页码：411 / 416

页数：6

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