Crystallization and preliminary X-ray diffraction analysis of 5,10-methylenetetrahydrofolate dehydrogenase/cyclohydrolase from Thermoplasma acidophilum DSM 1728

Jae Hee Kim, Min Woo Sung, Eun Hye Lee, Ki Hyun Nam, Kwang Yeon Hwang

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

The methylenetetrahydrofolate dehydrogenase/cyclohydrolase (MTHFDC) from the thermoacidophilic archaeon Thermoplasma acidophilum is a 30.6 kDa molecular-mass enzyme that sequentially catalyzes the conversion of formyltetrahydrofolate to methylenetetrahydrofolate, with a preference for NADP as a cofactor, rather than NAD. In order to elucidate the functional and structural features of MTHFDC from archaeons at a molecular level, it was overexpressed in Escherichia coli and crystallized in the presence of its cofactor, NADP, at 295 K using polyethylene glycol (PEG) 4000 as a precipitant. The crystal is a member of the monoclinic space group P211, with the following unit cell parameters: a=66.333 Å, b=52.868 Å, c=86.099 Å, and β=97.570°, and diffracts to a resolution of at least 2.40 Å at the synchrotron. Assuming a dimer in the crystallographic asymmetric unit, the calculated Matthews parameter (VM) was 2.44 Å3/Da and the solvent content was 49.7%.

Original languageEnglish
Pages (from-to)283-286
Number of pages4
JournalJournal of Microbiology and Biotechnology
Volume18
Issue number2
Publication statusPublished - 2008 Feb 28

Fingerprint

Methylenetetrahydrofolate Dehydrogenase (NADP)
Thermoplasma
Archaea
Crystallization
NADP
X-Ray Diffraction
X ray diffraction analysis
Formyltetrahydrofolates
Synchrotrons
Molecular mass
Dimers
NAD
Escherichia coli
Polyethylene glycols
Enzymes
Crystals
Oxidoreductases

Keywords

  • Crystallization
  • Cyclohydrolase
  • Dehydrogenase
  • Thermoplasma acidophilum

ASJC Scopus subject areas

  • Biotechnology
  • Applied Microbiology and Biotechnology
  • Microbiology
  • Bioengineering

Cite this

Crystallization and preliminary X-ray diffraction analysis of 5,10-methylenetetrahydrofolate dehydrogenase/cyclohydrolase from Thermoplasma acidophilum DSM 1728. / Kim, Jae Hee; Sung, Min Woo; Lee, Eun Hye; Nam, Ki Hyun; Hwang, Kwang Yeon.

In: Journal of Microbiology and Biotechnology, Vol. 18, No. 2, 28.02.2008, p. 283-286.

Research output: Contribution to journalArticle

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