Purification and some properties of cobalamin-dependent methionine synthase from rat liver

Kazuhiro Yamada, Takamasa Tobimatsu, Tetsunori Kawata, Masahiro Wada, Akio Maekawa, Tetsuo Toraya

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Cobalamin-dependent methionine synthase was purified from rat liver. The enzyme activity was separated into two peaks upon Mono-Q column chromatography. Peaks I and II of the enzyme, eluted in this order, were purified 18,000- and 44,000-fold in overall yields of 0.7 and 1.8%, respectively. Peak II methionine synthase, the major fraction, was homogeneous as judged by SDS-polyacrylamide gel electrophoresis. The enzyme was a large monomeric protein with an apparent molecular weight of 143,000 Da. Interconversion of the enzyme between the two peaks was not observed during purification procedures. The enzyme required S-adenosylmethionine and a reducing system for activity. Apparent Km values of the peak II enzyme for 5-methyltetrahydrofolate and homocysteine were 75 and 1.7μM, respectively.

Original languageEnglish
Pages (from-to)177-186
Number of pages10
JournalJournal of Nutritional Science and Vitaminology
Volume43
Issue number2
Publication statusPublished - 1997

Fingerprint

5-Methyltetrahydrofolate-Homocysteine S-Methyltransferase
methionine synthase
Vitamin B 12
liver
Liver
rats
Enzymes
enzymes
S-adenosylmethionine
homocysteine
S-Adenosylmethionine
Homocysteine
polyacrylamide gel electrophoresis
chromatography
Chromatography
Polyacrylamide Gel Electrophoresis
enzyme activity
molecular weight
Molecular Weight
proteins

Keywords

  • 5-methyltetrahydrofolate- homocysteine methyltransferase
  • cobalamin
  • methionine synthase
  • rat liver
  • S-adenosylmethionine
  • vitamin B

ASJC Scopus subject areas

  • Medicine (miscellaneous)
  • Food Science

Cite this

Purification and some properties of cobalamin-dependent methionine synthase from rat liver. / Yamada, Kazuhiro; Tobimatsu, Takamasa; Kawata, Tetsunori; Wada, Masahiro; Maekawa, Akio; Toraya, Tetsuo.

In: Journal of Nutritional Science and Vitaminology, Vol. 43, No. 2, 1997, p. 177-186.

Research output: Contribution to journalArticle

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AU - Toraya, Tetsuo

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