Enhanced Expression of an Iron–Sulfur Protein Slr0351 of Synechocystis sp. PCC 6803 in E. coli by Truncating the Transmembrane Region
- 作者: Ma Q.1,2, Lei H.1,2, Yan R.1, Zhou M.3
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隶属关系:
- Biological Scientific and Technical College
- Key Laboratory of Biologic Resources Protection and Utilization of Hubei Province
- State Key Laboratory of Agricultural Microbiology
- 期: 卷 44, 编号 2 (2018)
- 页面: 188-192
- 栏目: Article
- URL: https://journals.rcsi.science/1068-1620/article/view/228866
- DOI: https://doi.org/10.1134/S1068162018020073
- ID: 228866
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详细
Iron–sulfur proteins are important components of the photosynthetic electron transport complex in thylakoid membrane of cyanobacteria. The aim of this study was to enhance the expression of a putative iron–sulfur protein Slr0351 in E. coli by truncating the transmembrane region and to explore the iron–sulfur cluster binding properties of Slr0351. We truncated the N-terminal transmembrane region of Slr0351 (sample Slr0351_tr), which resulted in higher yield and higher solubility of Slr0351_tr expressed in E. coli BL21 (DE3) without supplementation of Fe2+ and S2– in LB media, compared with the full-length Slr0351. Using affinity chromatography under anaerobic conditions, we obtained purified Slr0351 and Slr0351_tr. Unlike full-length Slr0351, Slr0351_tr was of brown color and showed distinct absorption peak at 460 nm, which is characteristic of Fe/S cluster. To identify the binding cysteine site of Fe/S cluster in Slr0351_tr, we obtained four mutants of Slr0351_tr via site-directed mutagenesis. The binding sites were identified as C140, C141, and C288 based on disappearance of the absorbance at 460 nm characteristic of the mutants. These results confirm that Slr0351 is an iron–sulfur protein.
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作者简介
Qiong Ma
Biological Scientific and Technical College; Key Laboratory of Biologic Resources Protection and Utilization of Hubei Province
编辑信件的主要联系方式.
Email: maqiong110@126.com
中国, Enshi, 445000; Enshi, 445000
Hong-ling Lei
Biological Scientific and Technical College; Key Laboratory of Biologic Resources Protection and Utilization of Hubei Province
Email: maqiong110@126.com
中国, Enshi, 445000; Enshi, 445000
Rong Yan
Biological Scientific and Technical College
Email: maqiong110@126.com
中国, Enshi, 445000
Ming Zhou
State Key Laboratory of Agricultural Microbiology
Email: maqiong110@126.com
中国, Wuhan, 430070