Open AccessExpression, purification and crystallization of the FP domain of the human F‐box protein Fbxo7
Author(s) -
Shang Jinsai,
Wang Guan,
Yang Yang,
Huang Xiaolan,
Du Zhihua
Publication year - 2013
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309113023154
Subject(s) - ubiquitin ligase , ubiquitin , crystallization , proteasome , f box protein , chemistry , dna ligase , crystallography , protein domain , protein crystallization , biochemistry , microbiology and biotechnology , biology , dna , gene , organic chemistry
Fbxo7 is a conserved protein in higher eukaryotes that belongs to the F‐box protein family. Fbxo7 is the substrate‐recognition component of the SCF Fbxo7 (Skp1‐Cul1‐Fbox protein) E3 ubiquitin ligase. Besides the F‐box motif, Fbxo7 also contains a C‐terminal proline‐rich region, an N‐terminal ubiquitin‐like domain and a novel FP (Fbxo7/PI31) domain preceding the F‐box motif. The FP domains of Fbxo7 and the PI31 proteasome inhibitor mediate interaction between the two proteins. For structure determination of the FP domain of Fxbo7, a protein construct (amino acids 181–335) corresponding to the FP domain was expressed, purified and crystallized. The native and selenomethionine‐labeled proteins crystallized in different crystal forms. Native and single‐wavelength anomalous dispersion data sets with diffraction to 2.1 and 2.0 Å resolution, respectively, have been collected and structure determination is in progress.