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Crystallization and preliminary X-ray crystallographic analysis of human FAF1 UBX domain
Wonchull Kang1, Hwa Young Shin, Jin Kuk Yang
1Department of Chemistry, Soongsil University, Seoul 156-743, Republic of Korea.
Abstract:
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.
Insights
Fas-associated factor 1 (FAF1) protein structure was determined to understand its role in regulating protein degradation. This research provides insights into FAF1
Area of Science:
- Molecular Biology
- Structural Biology
- Biochemistry
Background:
- Fas-associated factor 1 (FAF1) is a key protein regulating apoptosis and the ubiquitin-proteasome pathway.
- FAF1 interacts with p97/VCP, a chaperone involved in proteasomal degradation, via its UBX domain.
- Understanding this interaction is crucial for elucidating FAF1's role in protein degradation modulation.
Purpose of the Study:
- To determine the crystal structure of the unbound FAF1 UBX domain.
- To provide a structural basis for FAF1's function in modulating p97/VCP activity.
- To facilitate comparison with the FAF1 UBX-p97/VCP N domain complex structure.
Main Methods:
- Crystallographic analysis of the unbound FAF1 UBX domain.
- X-ray diffraction data collection to 3.00 A resolution.
- Space group determination (F4(1)32) and unit-cell parameter calculation (a = b = c = 176.40 A).
Main Results:
- Crystals of the unbound FAF1 UBX domain were obtained.
- The crystal structure belongs to space group F4(1)32.
- Molecular replacement confirmed two molecules in the asymmetric unit, consistent with estimated Matthews coefficient and solvent content.
Conclusions:
- The structural data of the unbound FAF1 UBX domain are available for further analysis.
- This structure serves as a basis for understanding FAF1's interaction with p97/VCP.
- Further studies can explore how FAF1 binding affects p97/VCP function in proteasomal degradation.
