Crystal Structure of the C-Terminal Cytoplasmic Domain of Non-Structural Protein 4 from Mouse Hepatitis Virus A59

Xu, Xiaoling; Lou, Zhiyong; Ma, Yanlin; Chen, Xuehui; Yang, Zhangsheng; Tong, Xiaohang; Zhao, Qi; Xu, Yuanyuan; Deng, Hongyu; Bartlam, Mark; Rao, Zihe

The crystals were cryoprotected in a solution containing the reservoir solution with 15% glycerol and flash-cooled with liquid nitrogen. A 2.0 Ã… resolution data set of native nsp4C was collected at 100 K on beamline BL-5A of the Photon Factory in Japan using an ADSC Q315 detector. The crystal form belongs to the cubic space group C2 (a = 86.86, b = 52.55 Ã… , c = 53.87 Ã… , a = c = 90u, b = 116u) with two molecules in one asymmetric unit and a Matthews coefficient V M of 2.6 Ã… 3 Da 21 , corresponding to a solvent content of 57% [41] . The Se-Met derivative data of nsp4C were collected at BSRF (Beijing Synchrotron Radiation Facility) at 2.4 Ã… resolution using a Mar CCD detector. The Se-Met crystals shared the same space group and similar unit cell parameters with the native crystals. Data for the C425S mutant were collected to 2.4 Ã… resolution on beamline BL-17A of the Photon Factory in Japan using an ADSC Q270 detector. The crystal form belongs to the primitive tetragonal space group P4 1 2 1 2 (a = b = 54.79 Ã… , c = 97.55 Ã… , a = b = c = 90u). Processing of all diffraction images and scaling of the integrated intensities were performed using the HKL2000 software package [42] .