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Expression, purification, and characterization of human cytosolic serine hydroxymethyltransferase.
Protein Expr Purif. 1995 Aug; 6(4):411-6.PE

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.

Authors+Show Affiliations

Department of Biochemistry and Molecular Biophysics, Virginia Commonwealth University, Richmond 23298-0614, USA.No affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, U.S. Gov't, P.H.S.

Language

eng

PubMed ID

8527925

Citation

Kruschwitz, H, et al. "Expression, Purification, and Characterization of Human Cytosolic Serine Hydroxymethyltransferase." Protein Expression and Purification, vol. 6, no. 4, 1995, pp. 411-6.
Kruschwitz H, Ren S, Di Salvo M, et al. Expression, purification, and characterization of human cytosolic serine hydroxymethyltransferase. Protein Expr Purif. 1995;6(4):411-6.
Kruschwitz, H., Ren, S., Di Salvo, M., & Schirch, V. (1995). Expression, purification, and characterization of human cytosolic serine hydroxymethyltransferase. Protein Expression and Purification, 6(4), 411-6.
Kruschwitz H, et al. Expression, Purification, and Characterization of Human Cytosolic Serine Hydroxymethyltransferase. Protein Expr Purif. 1995;6(4):411-6. PubMed PMID: 8527925.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Expression, purification, and characterization of human cytosolic serine hydroxymethyltransferase. AU - Kruschwitz,H, AU - Ren,S, AU - Di Salvo,M, AU - Schirch,V, PY - 1995/8/1/pubmed PY - 1995/8/1/medline PY - 1995/8/1/entrez SP - 411 EP - 6 JF - Protein expression and purification JO - Protein Expr. Purif. VL - 6 IS - 4 N2 - 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. SN - 1046-5928 UR - https://www.unboundmedicine.com/medline/citation/8527925/Expression_purification_and_characterization_of_human_cytosolic_serine_hydroxymethyltransferase_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S1046592885710558 DB - PRIME DP - Unbound Medicine ER -