Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Crystallization and Preliminary X-Ray Diffraction Analysis of 5,10-Methylenetetrahydrofolate Dehydrogenase/Cyclohydrolase from Thermoplasma acidophilum DSM 1728

  • Kim, Jae-Hee (Division of Biotechnology, College of Life Sciences and Biotechnology, Korea University) ;
  • Sung, Min-Woo (Division of Biotechnology, College of Life Sciences and Biotechnology, Korea University) ;
  • Lee, Eun-Hye (Division of Biotechnology, College of Life Sciences and Biotechnology, Korea University) ;
  • Nam, Ki-Hyun (Division of Biotechnology, College of Life Sciences and Biotechnology, Korea University) ;
  • Hwang, Kwang-Yeon (Division of Biotechnology, College of Life Sciences and Biotechnology, Korea University)
  • Published : 2008.02.29
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Abstract

The methylenetetrahydrofolate dehydrogenase/cyclohydrolase (MTHFDC) from the thermoacidophilic archaeon Thermoplasma acidophilum is a 30.6kDa molecular-mass enzyme that sequentially catalyzes the conversion of formyltetrahydrofollate 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 295K using polyethylene glycol (PEG) 4000 as a precipitant. The crystal is a member of the monoclinic space group $P2_1$, with the following unit cell parameters: $a=66.333{\AA},\;b=52.868{\AA},\;c=86.099{\AA},\;and\;{\beta}=97.570^{\circ}$, and diffracts to a resolution of at least $2.40{\AA}$ at the synchrotron. Assuming a dimer in the crystallographic asymmetric unit, the calculated Matthews parameter $(V_M)\;was\;2.44{\AA}^3/Da$ and the solvent content was 49.7%.

Keywords

References

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