Biosynthesis of fluorescent cyanobacterial allophycocyanin trimer in Escherichia coli
文献类型：期刊论文
| 作者 | Liu, Shaofang1,2; Chen, Yingjie1,2; Lu, Yandu2,3; Chen, Huaxin1; Li, Fuchao1; Qin, Song1 |
| 刊名 | PHOTOSYNTHESIS RESEARCH
 |
| 出版日期 | 2010-08-01 |
| 卷号 | 105期号:2页码:135-142 |
| 关键词 | Allophycocyanin Trimer Assembly Recombinant Protein Allophycocyanin Monomer Fluorescence Escherichia Coli |
| ISSN号 | 0166-8595 |
| DOI | 10.1007/s11120-010-9574-4 |
| 文献子类 | Article |
| 英文摘要 | Allophycocyanin (APC), a cyanobacterial photosynthetic phycobiliprotein, functions in energy transfer as a light-harvesting protein. One of the prominent spectroscopic characteristics of APC is a strong red-shift in the absorption and emission maxima when monomers are assembled into a trimer. Previously, holo-APC alpha and beta subunits (holo-ApcA and ApcB) were successfully synthesized in Escherichia coli. In this study, both holo-subunits from Synechocystis sp. PCC 6803 were co-expressed in E. coli, and found to self-assemble into trimers. The recombinant APC trimer was purified by metal affinity and size-exclusion chromatography, and had a native structure identical to native APC, as determined by characteristic spectroscopic measurements, fluorescence quantum yield, tryptic digestion analysis, and molecular weight measurements. Combined with results from a study in which only the monomer was formed, our results indicate that bilin synthesis and the subsequent attachment to apo-subunits are important for the successful assembly of APC trimers. This is the first study to report on the assembly of recombinant ApcA and ApcB into a trimer with native structure. Our study provides a promising method for producing better fluorescent tags, as well as a method to facilitate the genetic analysis of APC trimer assembly and biological function.; Allophycocyanin (APC), a cyanobacterial photosynthetic phycobiliprotein, functions in energy transfer as a light-harvesting protein. One of the prominent spectroscopic characteristics of APC is a strong red-shift in the absorption and emission maxima when monomers are assembled into a trimer. Previously, holo-APC alpha and beta subunits (holo-ApcA and ApcB) were successfully synthesized in Escherichia coli. In this study, both holo-subunits from Synechocystis sp. PCC 6803 were co-expressed in E. coli, and found to self-assemble into trimers. The recombinant APC trimer was purified by metal affinity and size-exclusion chromatography, and had a native structure identical to native APC, as determined by characteristic spectroscopic measurements, fluorescence quantum yield, tryptic digestion analysis, and molecular weight measurements. Combined with results from a study in which only the monomer was formed, our results indicate that bilin synthesis and the subsequent attachment to apo-subunits are important for the successful assembly of APC trimers. This is the first study to report on the assembly of recombinant ApcA and ApcB into a trimer with native structure. Our study provides a promising method for producing better fluorescent tags, as well as a method to facilitate the genetic analysis of APC trimer assembly and biological function. |
| 学科主题 | Plant Sciences |
| URL标识 | 查看原文 |
| 语种 | 英语 |
| WOS记录号 | WOS:000280921200006 |
| 公开日期 | 2010-12-24 |
| 源URL | [http://ir.qdio.ac.cn/handle/337002/5793]  |
| 专题 | 海洋研究所_实验海洋生物学重点实验室 |
| 作者单位 | 1.Chinese Acad Sci, Key Lab Expt Marine Biol, Inst Oceanol, Qingdao 266071, Peoples R China 2.Chinese Acad Sci, Grad Sch, Beijing 100049, Peoples R China 3.Chinese Acad Sci, Yantai Inst Coastal Zone Res, Yantai 264003, Peoples R China |
| 推荐引用方式 GB/T 7714 | Liu, Shaofang,Chen, Yingjie,Lu, Yandu,et al. Biosynthesis of fluorescent cyanobacterial allophycocyanin trimer in Escherichia coli[J]. PHOTOSYNTHESIS RESEARCH,2010,105(2):135-142. |
| APA | Liu, Shaofang,Chen, Yingjie,Lu, Yandu,Chen, Huaxin,Li, Fuchao,&Qin, Song.(2010).Biosynthesis of fluorescent cyanobacterial allophycocyanin trimer in Escherichia coli.PHOTOSYNTHESIS RESEARCH,105(2),135-142. |
| MLA | Liu, Shaofang,et al."Biosynthesis of fluorescent cyanobacterial allophycocyanin trimer in Escherichia coli".PHOTOSYNTHESIS RESEARCH 105.2(2010):135-142. |
入库方式： OAI收割
来源：海洋研究所
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