Abstract
Human ankyrin repeat and suppressor of cytokine signaling box protein 9 (hASB9), a subunit of an Elongin Ccullin- SOCS box (ECS) E3 ubiquitin ligase complex, is believed to be involved in specific substrate-recognition for ubiquitination and degradation. In fact, this specific substrate-recognition is determined by the ankyrin repeats of hASB9 protein. Here, we have cloned and overexpressed the hASB9-2, the splice variant of hASB9 with only one ankyrin repeat domain, as a 6His-tagged recombinant protein in Escherichia coli. The purified hASB9-2 protein was crystallized by the hanging-drop vapor-diffusion technique and diffracted to 2.2Å resolution. The data showed that the cubic hASB9-2 crystal belongs to space group P4332 with unit-cell parameters (a=b=c=129.25Å, α=β=γ=90°). An asymmetric unit in the crystal was assumed to contain one protein molecule giving the Matthews Coefficient factor of 2.81 and the solvent content of 56.3%.
Keywords: hASB9, hASB9-2, ankyrin repeat, crystallization, preliminary X-ray analysis
Protein & Peptide Letters
Title: Crystallization and Preliminary X-Ray Analysis of the Splice Variant of Human Ankyrin Repeat and Suppressor of Cytokine Signaling Box Protein 9 (hASB9-2)
Volume: 16 Issue: 3
Author(s): Xiangwei Fei, Yong Zhang, Xing Gu, Rui Qiu, Yumin Mao and Chaoneng Ji
Affiliation:
Keywords: hASB9, hASB9-2, ankyrin repeat, crystallization, preliminary X-ray analysis
Abstract: Human ankyrin repeat and suppressor of cytokine signaling box protein 9 (hASB9), a subunit of an Elongin Ccullin- SOCS box (ECS) E3 ubiquitin ligase complex, is believed to be involved in specific substrate-recognition for ubiquitination and degradation. In fact, this specific substrate-recognition is determined by the ankyrin repeats of hASB9 protein. Here, we have cloned and overexpressed the hASB9-2, the splice variant of hASB9 with only one ankyrin repeat domain, as a 6His-tagged recombinant protein in Escherichia coli. The purified hASB9-2 protein was crystallized by the hanging-drop vapor-diffusion technique and diffracted to 2.2Å resolution. The data showed that the cubic hASB9-2 crystal belongs to space group P4332 with unit-cell parameters (a=b=c=129.25Å, α=β=γ=90°). An asymmetric unit in the crystal was assumed to contain one protein molecule giving the Matthews Coefficient factor of 2.81 and the solvent content of 56.3%.
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Fei Xiangwei, Zhang Yong, Gu Xing, Qiu Rui, Mao Yumin and Ji Chaoneng, Crystallization and Preliminary X-Ray Analysis of the Splice Variant of Human Ankyrin Repeat and Suppressor of Cytokine Signaling Box Protein 9 (hASB9-2), Protein & Peptide Letters 2009; 16 (3) . https://dx.doi.org/10.2174/092986609787601688
DOI https://dx.doi.org/10.2174/092986609787601688 |
Print ISSN 0929-8665 |
Publisher Name Bentham Science Publisher |
Online ISSN 1875-5305 |
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