title: Crystallization and preliminary X-ray crystallographic analysis of the hexameric human p97/VCP ND1 fragment in complex with the UBX domain of human FAF1.
abstract: 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 I222, 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.
authors.affiliation: Department of Chemistry, College of Natural Sciences, Soongsil University, Seoul 156-743, Republic of Korea.
authors.forename: Wonchull
authors.initials: W
authors.lastname: Kang
authors.forename: Jin Kuk
authors.initials: JK
authors.lastname: Yang
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000737:chemistry* / Q000378:metabolism; D000251:Adenosine Triphosphatases / Q000737:chemistry* / Q000378:metabolism; D051017:Apoptosis Regulatory Proteins; D018797:Cell Cycle Proteins / Q000737:chemistry* / Q000378:metabolism; D018360:Crystallography, X-Ray; D006801:Humans; D055503:Protein Multimerization; D000074405:Valosin Containing Protein
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D051017:Apoptosis Regulatory Proteins; D018797:Cell Cycle Proteins; C490780:FAF1 protein, human; D000251:Adenosine Triphosphatases; C000614178:VCP protein, human; D000074405:Valosin Containing Protein
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