Open AccessCrystallization and preliminary X‐ray crystallographic analysis of the hexameric human p97/VCP ND1 fragment in complex with the UBX domain of human FAF1
Author(s) -
Kang Wonchull,
Yang Jin Kuk
Publication year - 2011
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309111031794
Subject(s) - crystallography , crystallization , ultrabithorax , mutant , materials science , chemistry , biochemistry , homeotic gene , organic chemistry , gene
The UBX domain of Fas‐associated factor 1 (FAF1) binds to the N domain of p97/VCP, a multi‐functional hexameric ATPase, and FAF1 thus inhibits the proteasome‐mediated protein‐degradation process assisted by p97/VCP. Here, crystallization of the hexameric p97/VCP ND1 fragment in complex with the FAF1 UBX domain is reported. Wild‐type p97/VCP ND1 in complex with FAF1 UBX crystallized into very thin sheet‐shaped crystals which turned out to be of poor diffraction quality. Therefore, in order to acquire a better diffraction‐quality crystal, three mutants of p97/VCP ND1 were generated based on the surface‐entropy reduction method. Of these, a triple mutant was the most successful in producing diffraction‐quality crystals suitable for subsequent structural analysis. X‐ray data were collected to 3.60 Å resolution and the crystals belonged to space group I 222, with unit‐cell parameters a = 166.28, b = 170.04, c = 255.99 Å. The Matthews coefficient and solvent content were estimated to be 5.78 Å 3 Da −1 and 78.72%, respectively.