Open AccessCrystallization and preliminary X‐ray characterization of the Skp1–Fbg3 complex
Author(s) -
Kumanomidou Taichi,
Nakagawa Tomomi,
Mizushima Tsunehiro,
Suzuki Atsuo,
Tokunaga Fuminori,
Iwai Kazuhiro,
Yoshida Yukiko,
Tanaka Keiji,
Yamane Takashi
Publication year - 2010
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309109050581
Subject(s) - crystallization , characterization (materials science) , materials science , x ray , skp1 , crystallography , chemistry , chemical engineering , nanotechnology , physics , optics , engineering , biochemistry , ubiquitin , gene , ubiquitin ligase
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 P 2 1 2 1 2 1 , with unit‐cell parameters a = 34.1, b = 76.6, c = 193.9 Å and one molecule per asymmetric unit.