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Biosynthesis of fluorescent cyanobacterial allophycocyanin trimer in Escherichia coli
Liu, Shaofang1,2; Chen, Yingjie1,2; Lu, Yandu2,3; Chen, Huaxin1; Li, Fuchao1; Qin, Song1
2010-08-01
Source PublicationPHOTOSYNTHESIS RESEARCH
ISSN0166-8595
Volume105Issue:2Pages:135-142
Contribution Rank[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
AbstractAllophycocyanin (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.
Corresponding AuthorQin, S, Chinese Acad Sci, Key Lab Expt Marine Biol, Inst Oceanol, Qingdao 266071, Peoples R China
KeywordAllophycocyanin Trimer Assembly Recombinant Protein Allophycocyanin Monomer Fluorescence Escherichia Coli
Department生物资源实验室 
Subject AreaPlant Sciences
Funding OrganizationChinese 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]
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Indexed BySCI
WOS KeywordLIGHT-HARVESTING PROTEINS ; SYNECHOCOCCUS SP PCC-7002 ; HOLO-ALPHA SUBUNIT ; C-PHYCOCYANIN ; MASTIGOCLADUS-LAMINOSUS ; HETEROLOGOUS HOST ; BETA-PHYCOCYANIN ; ENERGY-TRANSFER ; CPCS-I ; PHYCOBILIPROTEINS
SubtypeArticle
Language英语
WOS Research AreaPlant Sciences
WOS IDWOS:000280921200006
Citation statistics
Cited Times:17[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://ir.yic.ac.cn/handle/133337/3749
Collection海岸带生物学与生物资源保护实验室
Affiliation1.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
Recommended Citation
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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