Purification and crystallization of the catalytic PRONE domain of RopGEF8 and its complex with Rop4 from Arabidopsis thaliana.
The PRONE domain of the guanine nucleotide exchange factor RopGEF8 (PRONE8) was purified and crystallized free and in complex with the Rho-family protein Rop4 using the hanging-drop vapour-diffusion method. PRONE8 crystals were obtained using NaCl as precipitating agent and belong to the hexagonal space group P6(5)22. Native and anomalous data sets were collected using synchrotron radiation at 100 K to 2.2 and 2.8 A resolution, respectively. Crystals of the Rop4-PRONE8 complex belonging to space group P6(3) were obtained using Tacsimate and PEG 3350 as precipitating agents and diffracted to 3.1 A resolution.
Thomas
Christoph
C
Department of Structural Biology, Max Planck Institute of Molecular Physiology, Otto-Hahn-Strasse 11, 44227 Dortmund, Germany.
Weyand
Michael
M
Wittinghofer
Alfred
A
Berken
Antje
A
D029681:Arabidopsis Proteins / Q000737:chemistry* / Q000302:isolation & purification; D020134:Catalytic Domain; D003460:Crystallization / Q000379:methods; D019204:GTP-Binding Proteins / Q000737:chemistry* / Q000302:isolation & purification; D020662:Guanine Nucleotide Exchange Factors / Q000737:chemistry* / Q000302:isolation & purification; D011485:Protein Binding; D012997:Solvents; D014961:X-Ray Diffraction
D029681:Arabidopsis Proteins; D020662:Guanine Nucleotide Exchange Factors; D012997:Solvents; D019204:GTP-Binding Proteins; C511948:Rop4 protein, Arabidopsis