Expression, purification and preliminary X-ray analysis of the BRCT domain from Rhp9/Crb2.
The BRCT domain from Rhp9 (a Schizosaccharomyces pombe DNA-damage checkpoint protein) has been expressed, purified and crystallized. Overexpression in bacterial cells was achieved by minimizing aeration during host cell growth. A robotic screen was used to determine the solubility parameters; concentration of the protein was achieved by exploiting this information. Single crystals suitable for X-ray analysis were obtained in two forms by vapour diffusion (trigonal, unit-cell parameters a = b = 228.04, c = 70.42 A, and tetragonal, P4/m Laue group symmetry, unit-cell parameters a = b = 72.3, c = 91.1 A).
Hinks
J A
JA
CR-UK DNA Repair Enzyme Group, Institute of Cancer Research, 237 Fulham Road, South Kensington, London SW3 6JB, England.
Roe
M
M
Ho
J C Y
JC
Watts
F Z
FZ
Phelan
J
J
McAllister
M
M
Pearl
L H
LH
D001331:Automation; D018797:Cell Cycle Proteins / Q000737:chemistry*; D003001:Cloning, Molecular / Q000379:methods; D003460:Crystallization / Q000379:methods; D018360:Crystallography, X-Ray; D009687:Nuclear Proteins / Q000737:chemistry*; D017434:Protein Structure, Tertiary; D029702:Schizosaccharomyces pombe Proteins / Q000737:chemistry*; D012965:Sodium Chloride; D012995:Solubility
D018797:Cell Cycle Proteins; C109867:Crb2 protein, S pombe; D009687:Nuclear Proteins; D029702:Schizosaccharomyces pombe Proteins; D012965:Sodium Chloride