Kang Wonchull, Shin Hwa Young, Yang Jin Kuk
Department of Chemistry, Soongsil University, Seoul 156-743, Republic of Korea.
Acta Crystallogr Sect F Struct Biol Cryst Commun. 2010 Feb 1;66(Pt 2):211-3. doi: 10.1107/S1744309110001077. Epub 2010 Jan 28.
Fas-associated factor 1 (FAF1) is a multifunctional pro-apoptotic protein that is involved in Fas-mediated apoptosis, NF-kappaB signalling and the ubiquitin-proteasome pathway. In the ubiquitin-proteasome pathway, FAF1 binds to the N domain of p97/VCP, a molecular chaperone that acts in complex with the proteasome, through its C-terminal UBX domain and inhibits the proteasomal protein-degradation process. In an effort to elucidate the structural basis of the function of FAF1 in modulating p97/VCP activity related to proteasomal protein degradation, crystallographic analysis of the FAF1 UBX domain and the p97/VCP N domain was initiated. Following the recently reported crystallization of the FAF1 UBX domain bound to the p97/VCP N domain, the unbound FAF1 UBX domain was also crystallized for purposes of structural comparison. X-ray data were collected to 3.00 A resolution and the crystals belonged to space group F4(1)32, with unit-cell parameters a = b = c = 176.40 A. The Matthews coefficient and solvent content were estimated to be 3.04 A(3) Da(-1) and 59.5%, respectively, assuming that the asymmetric unit contained two molecules of the UBX domain, which was subsequently confirmed by molecular-replacement calculations.
Fas相关因子1（FAF1）是一种多功能促凋亡蛋白，参与Fas介导的凋亡、核因子κB信号传导和泛素-蛋白酶体途径。在泛素-蛋白酶体途径中，FAF1通过其C末端的UBX结构域与p97/VCP的N结构域结合，p97/VCP是一种与蛋白酶体协同作用的分子伴侣，可抑制蛋白酶体蛋白降解过程。为了阐明FAF1在调节与蛋白酶体蛋白降解相关的p97/VCP活性中的功能结构基础，我们开始对FAF1的UBX结构域和p97/VCP的N结构域进行晶体学分析。继最近报道了与p97/VCP N结构域结合的FAF1 UBX结构域的晶体结构后，为了进行结构比较，还对未结合的FAF1 UBX结构域进行了结晶。收集了分辨率为3.00 Å的X射线数据，晶体属于空间群F4(1)32，晶胞参数a = b = c = 176.40 Å。假设不对称单元包含两个UBX结构域分子，马修斯系数和溶剂含量分别估计为3.04 Å3 Da-1和59.5%，随后通过分子置换计算得到证实。
