Crystallization and preliminary X-ray diffraction study of an active-site mutant of pro-Tk-subtilisin from a hyperthermophilic archaeon.
Crystallization of and preliminary crystallographic studies on an active-site mutant of pro-Tk-subtilisin from the hyperthermophilic archaeon Thermococcus kodakaraensis were performed. The crystal was grown at 277 K by the sitting-drop vapour-diffusion method. Native X-ray diffraction data were collected to 2.3 A resolution using synchrotron radiation from station BL41XU at SPring-8. The crystal belongs to the orthorhombic space group I222, with unit-cell parameters a = 92.69, b = 121.78, c = 77.53 A. Assuming the presence of one molecule per asymmetric unit, the Matthews coefficient V(M) was calculated to be 2.6 A(3) Da(-1) and the solvent content was 53.1%.
Tanaka
Shun-ichi
S
Department of Material and Life Science, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita, Osaka 565-0871, Japan.
Saito
Kenji
K
Chon
Hyongi
H
Matsumura
Hiroyoshi
H
Koga
Yuichi
Y
Takano
Kazufumi
K
Kanaya
Shigenori
S
D019843:Archaeal Proteins / Q000737:chemistry* / Q000235:genetics*; D001665:Binding Sites / Q000235:genetics; D003460:Crystallization; D004792:Enzyme Precursors / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D009154:Mutation; D010446:Peptide Fragments / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D013381:Subtilisins / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D019712:Thermococcus / Q000737:chemistry* / Q000235:genetics*; D014961:X-Ray Diffraction / Q000379:methods
D019843:Archaeal Proteins; D004792:Enzyme Precursors; D010446:Peptide Fragments; C057107:prosubtilisin; D013381:Subtilisins