| Biosynthesis of fluorescent cyanobacterial allophycocyanin trimer in Escherichia coli | |
Liu, Shaofang1,2; Chen, Yingjie1,2; Lu, Yandu2,3; Chen, Huaxin1; Li, Fuchao1; Qin, Song1
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| 发表期刊 | PHOTOSYNTHESIS RESEARCH
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| ISSN | 0166-8595 |
| 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 |
| 产权排序 | [Liu, Shaofang; Chen, Yingjie; Chen, Huaxin; Li, Fuchao; Qin, Song] Chinese Acad Sci, Key Lab Expt Marine Biol, Inst Oceanol, Qingdao 266071, Peoples R China; [Liu, Shaofang; Chen, Yingjie; Lu, Yandu] Chinese Acad Sci, Grad Sch, Beijing 100049, Peoples R China; [Lu, Yandu] Chinese Acad Sci, Yantai Inst Coastal Zone Res, Yantai 264003, Peoples R China |
| 通讯作者 | Qin, S, Chinese Acad Sci, Key Lab Expt Marine Biol, Inst Oceanol, Qingdao 266071, Peoples R China |
| 作者部门 | 生物资源实验室 |
| 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. |
| 文章类型 | Article |
| 资助机构 | Chinese Academy of Sciences [KZCX2-YW-209, KZCX2-YW-216]; Hi-Tech Research and Development Program of China [2006 AA090303]; Science & Technology Project of Qingdao [06-2-2-12-JCH, 09-1-3-59-JCH] |
| 收录类别 | SCI |
| 语种 | 英语 |
| 关键词[WOS] | LIGHT-HARVESTING PROTEINS ; SYNECHOCOCCUS SP PCC-7002 ; HOLO-ALPHA SUBUNIT ; C-PHYCOCYANIN ; MASTIGOCLADUS-LAMINOSUS ; HETEROLOGOUS HOST ; BETA-PHYCOCYANIN ; ENERGY-TRANSFER ; CPCS-I ; PHYCOBILIPROTEINS |
| 研究领域[WOS] | Plant Sciences |
| WOS记录号 | WOS:000280921200006 |
| 引用统计 | |
| 文献类型 | 期刊论文 |
| 条目标识符 | http://ir.yic.ac.cn/handle/133337/3749 |
| 专题 | 海岸带生物学与生物资源利用重点实验室_海岸带生物学与生物资源保护实验室 |
| 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. |
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