Crystallization and preliminary X-ray analysis of 5′-methylthioribose kinase from Bacillus subtilis and Arabidopsis thaliana

Shao Yang Ku, Patrick Yip, Kenneth A. Cornell, Michael Riscoe, P. L. Howell

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

Recombinant Bacillus subtilis 5′-methylthioribose (MTR) kinase has been expressed, purified and subsequently crystallized using the hanging-drop vapor-diffusion technique. With PEG 2000MME as the precipitant, two different crystal forms have been grown in the absence and presence of the detergent CHAPS. These crystals belong to space groups P21212 1 (unit-cell parameters a = 193.7, b = 83.2, c = 51.6 Å) and P21212 (unit-cell parameters a = 213.8, b = 83.2, c = 51.5 Å), respectively. The crystals grown in the presence of CHAPS diffract to 2.2 Å resolution at Station X8C, National Synchrotron Light Source (NSLS). For both crystal forms, the presence of two monomers per asymmetric unit is predicted (Matthews coefficient VM = 2.29 and 2.52 Å3 Da-1, respectively). Recombinant C-terminally histidine-tagged Arabidopsis thaliana MTR kinase has also been expressed, purified and refolded into its active form. Rod-shaped crystals of this protein were grown from PEG 8000 using the hanging-drop vapor-diffusion technique. These crystals exhibit the symmetry of space group C2 (unit-cell parameters a = 162.3, b = 83.3, c = 91.0 Å, β = 117.8°) and diffract to 1.9 Å resolution at Station X8C, NSLS. Two monomers are estimated to be present in the asymmetric unit (VM = 2.82 Å3 Da-1).

Original languageEnglish (US)
Pages (from-to)116-119
Number of pages4
JournalActa Crystallographica Section D: Biological Crystallography
Volume60
Issue number1
DOIs
StatePublished - Jan 2004

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Bacillus
X ray analysis
Bacilli
Crystallization
Bacillus subtilis
Arabidopsis
Synchrotrons
X-Rays
crystallization
Crystals
crystals
Light
x rays
Histidine
Detergents
Light sources
light sources
synchrotrons
monomers
stations

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry
  • Biophysics
  • Condensed Matter Physics
  • Structural Biology

Cite this

Crystallization and preliminary X-ray analysis of 5′-methylthioribose kinase from Bacillus subtilis and Arabidopsis thaliana. / Ku, Shao Yang; Yip, Patrick; Cornell, Kenneth A.; Riscoe, Michael; Howell, P. L.

In: Acta Crystallographica Section D: Biological Crystallography, Vol. 60, No. 1, 01.2004, p. 116-119.

Research output: Contribution to journalArticle

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