Crystallization and preliminary X-ray crystallographic analysis of the N domain of p97/VCP in complex with the UBX domain of FAF1

• Published in: Acta crystallographica. Section F, Structural biology and crystallization communications Structural biology and crystallization communications, 2010-01, Vol.66 (1), p.41-43
• Main Authors: Shin, Hwa Young, Kang, Wonchull, Lee, Sang Yoon, Yang, Jin Kuk
• Format: Article
• Language: English
• Subjects: Adaptor Proteins, Signal Transducing - chemistry; Adenosine Triphosphatases - chemistry; Apoptosis Regulatory Proteins; Cell Cycle Proteins - chemistry; Crystallization; Crystallization Communications; Crystallography, X-Ray; FAF1; Humans; p97; Protein Structure, Tertiary; UBX; Valosin Containing Protein; VCP
• ISSN: 1744-3091; 1744-3091; 2053-230X
• DOI: 10.1107/S1744309109047691

p97/VCP is a multifunctional AAA+‐family ATPase that is involved in diverse cellular processes. p97/VCP directly interacts with various adaptors for activity in different biochemical contexts. Among these adaptors are p47 and Fas‐associated factor 1 (FAF1), which contain a common UBX domain through which they bind to the N domain of p97/VCP. In the ubiquitin–proteasome pathway, p97/VCP acts as a chaperone that presents client proteins to the proteasome for degradation, while FAF1 modulates the process by interacting with ubiquitinated client proteins and also with p97/VCP. In an effort to elucidate the structural details of the interaction between p97/VCP and FAF1, the p97/VCP N domain was crystallized in complex with the FAF1 UBX domain. X‐ray data were collected to 2.60 Å resolution and the crystals belonged to space group C2221, with unit‐cell parameters a = 58.24, b = 72.81, c = 132.93 Å. The Matthews coefficient and solvent content were estimated to be 2.39 Å3 Da−1 and 48.4%, respectively, assuming that the asymmetric unit contained p97/VCP N domain and FAF1 molecules in a 1:1 ratio, which was subsequently confirmed by molecular‐replacement calculations.