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Crystallization and preliminary X-ray characterization of the Skp1-Fbg3 complex.

ABSTRACT: 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.

SUBMITTER: Kumanomidou T

PROVIDER: S-EPMC2805547 | biostudies-literature | 2010 Jan

REPOSITORIES: biostudies-literature

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Crystallization and preliminary X-ray characterization of the Skp1-Fbg3 complex.

Kumanomidou Taichi T Nakagawa Tomomi T Mizushima Tsunehiro T Suzuki Atsuo A Tokunaga Fuminori F Iwai Kazuhiro K Yoshida Yukiko Y Tanaka Keiji K Yamane Takashi T

Acta crystallographica. Section F, Structural biology and crystallization communications 20091225 Pt 1

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 compl ...[more]

PMID: 20057081

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