Crystallization and preliminary X-ray characterization of the Skp1-Fbg3 complex.
F-box proteins are the substrate-recognition components of Skp1-Cullin1-F-box protein-Rbx1 (SCF) ubiquitin ligase complexes. Fbs1, an F-box protein, binds specifically to proteins modified with high-mannose oligosaccharides. Fbg3, another F-box protein, has 51% sequence identity to Fbs1. Although the residues that are necessary for binding to oligosaccharides are conserved between Fbs1 and Fbg3, Fbg3 does not bind glycoproteins. Skp1 and Fbg3 were co-expressed in Escherichia coli and their complex was purified to homogeneity and crystallized. Microseeding combined with the sandwiched hanging-drop technique improved the quality of the resulting crystals. The plate-shaped crystals belonged to space group P2(1)2(1)2(1), with unit-cell parameters a = 34.1, b = 76.6, c = 193.9 A and one molecule per asymmetric unit.
Kumanomidou
Taichi
T
Department of Biotechnology, Graduate School of Engineering, Nagoya University, Chikusa-ku, Nagoya 464-8603, Japan.
Nakagawa
Tomomi
T
Mizushima
Tsunehiro
T
Suzuki
Atsuo
A
Tokunaga
Fuminori
F
Iwai
Kazuhiro
K
Yoshida
Yukiko
Y
Tanaka
Keiji
K
Yamane
Takashi
T
D003460:Crystallization; D018360:Crystallography, X-Ray; D044783:F-Box Proteins / Q000737:chemistry*; D006801:Humans; D044843:SKP Cullin F-Box Protein Ligases / Q000737:chemistry*
D044783:F-Box Proteins; D044843:SKP Cullin F-Box Protein Ligases