Characterization of chlorophyll f synthase heterologously produced in Synechococcus sp. PCC 7002

被引:48
|
作者
Shen, Gaozhong [1 ]
Canniffe, Daniel P. [1 ,5 ]
Ho, Ming-Yang [1 ]
Kurashov, Vasily [1 ]
van der Est, Art [3 ]
Golbeck, John H. [1 ,2 ]
Bryant, Donald A. [1 ,4 ,6 ]
机构
[1] Penn State Univ, Dept Biochem & Mol Biol, University Pk, PA 16802 USA
[2] Penn State Univ, Dept Chem, University Pk, PA 16802 USA
[3] Brock Univ, Dept Chem, St Catharines, ON L2S 3A1, Canada
[4] Montana State Univ, Dept Chem & Biochem, Bozeman, MT 59717 USA
[5] Univ Sheffield, Dept Mol Biol & Biotechnol, Sheffield S10 2TN, S Yorkshire, England
[6] Penn State Univ, Dept Biochem & Mol Biol, Frear Lab S002, University Pk, PA 16802 USA
基金
美国国家科学基金会;
关键词
FaRLiP; Chlorophyll; Photosynthesis; Fischerella thermalis PCC 7521; Cyanobacteria; Photosystem I; FAR-RED LIGHT; PHOTOSYSTEM-II; PHOTOACCLIMATION FARLIP; ELECTRON-TRANSFER; WATER OXIDATION; CYANOBACTERIUM; OXYGEN; GENES; PHYCOBILIPROTEINS; BIOSYNTHESIS;
D O I
10.1007/s11120-018-00610-9
中图分类号
Q94 [植物学];
学科分类号
071001 ;
摘要
In diverse terrestrial cyanobacteria, Far-Red Light Photoacclimation (FaRLiP) promotes extensive remodeling of the photosynthetic apparatus, including photosystems (PS)I and PSII and the cores of phycobilisomes, and is accompanied by the concomitant biosynthesis of chlorophyll (Chl) d and Chl f. Chl f synthase, encoded by chlF, is a highly divergent paralog of psbA; heterologous expression of chlF from Chlorogloeopsis fritscii PCC 9212 led to the light-dependent production of Chl f in Synechococcus sp. PCC 7002 (Ho et al., Science 353, aaf9178 (2016)). In the studies reported here, expression of the chlF gene from Fischerella thermalis PCC 7521 in the heterologous system led to enhanced synthesis of Chl f. N-terminally [His](10)-tagged ChlF(7521) was purified and identified by immunoblotting and tryptic-peptide mass fingerprinting. As predicted from its sequence similarity to PsbA, ChlF bound Chl a and pheophytin a at a ratio of 3-4:1, bound -carotene and zeaxanthin, and was inhibited in vivo by 3-(3,4-dichlorophenyl)-1,1-dimethylurea. Cross-linking studies and the absence of copurifying proteins indicated that ChlF forms homodimers. Flash photolysis of ChlF produced a Chl a triplet that decayed with a lifetime (1/e) of 817 mu s and that could be attributed to intersystem crossing by EPR spectroscopy at 90K. When the chlF(7521) gene was expressed in a strain in which the psbD1 and psbD2 genes had been deleted, significantly more Chl f was produced, and Chl f levels could be further enhanced by specific growth-light conditions. Chl f synthesized in Synechococcus sp. PCC 7002 was inserted into trimeric PSI complexes.
引用
收藏
页码:77 / 92
页数:16
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