Crystallization and preliminary X-ray analysis of 5'-methylthioribose kinase from Bacillus subtilis and Arabidopsis thaliana.
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 P2(1)2(1)2(1) (unit-cell parameters a = 193.7, b = 83.2, c = 51.6 A) and P2(1)2(1)2 (unit-cell parameters a = 213.8, b = 83.2, c = 51.5 A), respectively. The crystals grown in the presence of CHAPS diffract to 2.2 A 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 V(M) = 2.29 and 2.52 A(3) 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 A, beta = 117.8 degrees ) and diffract to 1.9 A resolution at Station X8C, NSLS. Two monomers are estimated to be present in the asymmetric unit (V(M) = 2.82 A(3) Da(-1)).
Ku
Shao Yang
SY
Structural Biology and Biochemistry, Research Institute, Hospital for Sick Children, 555 University Avenue, Toronto, Ontario M5G 1X8, Canada.
Yip
Patrick
P
Cornell
Kenneth A
KA
Riscoe
Michael K
MK
Howell
P Lynne
PL
D017360:Arabidopsis / Q000201:enzymology* / Q000235:genetics; D001412:Bacillus subtilis / Q000201:enzymology* / Q000235:genetics; D002852:Chromatography, Ion Exchange; D003001:Cloning, Molecular; D003460:Crystallization; D018360:Crystallography, X-Ray; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D018744:DNA, Plant / Q000737:chemistry / Q000235:genetics; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000737:chemistry* / Q000235:genetics; D016133:Polymerase Chain Reaction; D017510:Protein Folding; D011994:Recombinant Proteins
D004269:DNA, Bacterial; D018744:DNA, Plant; D011994:Recombinant Proteins; D017853:Phosphotransferases (Alcohol Group Acceptor); C021787:5-methylthioribose kinase