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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Plants are still the major repository of biologically active substances. In the last two decades, however, natural peptides and proteins of plant origin have gained increasing attention due to their pharmacological activities over a variety of human illness, including those mediated by infections and parasitosis and those involving different cellular ...read more
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