Molecular mechanism of photoactivation and structural location of the cyanobacterial orange carotenoid protein.
Molecular mechanism of photoactivation and structural location of the cyanobacterial orange carotenoid protein.
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DOI：
10.1021/bi401539w
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发表时间：
2014-01-14
期刊：
影响因子：
2.9
通讯作者：
Blankenship, Robert E.
中科院分区：
文献类型：
--
作者：
Zhang, Hao;Liu, Haijun;Niedzwiedzki, Dariusz M.;Prado, Mindy;Jiang, Jing;Gross, Michael L.;Blankenship, Robert E.
橙类胡萝卜素蛋白（OCP）在蓝藻光合作用中具有与高等植物非光化学猝灭相似的光保护作用。在强光条件下，OCP与藻胆异构体（PBS）结合，减少向光系统的能量转移。保护周期从光诱导的OCP激活开始。关于这一过程的分子机制以及随后活性OCP向藻胆小体的募集的详细信息尚不清楚。本文报道了利用天然质谱（MS）和蛋白质交联相结合的方法，对蓝细菌Synechocystis sp. PCC 6803的OCP光激活进行了研究。我们证明了Native MS能够以完整的色素捕获OCP，并进一步揭示了OCP在光照下经历了二聚体-单体的转变。用天然质谱法观察了OCP的活化形态向无活性的暗形态的转变。此外，OCP和PBS的体外重构允许进行蛋白质化学交联实验。LC-MS/MS分析鉴定出OCP与PBS核心组分之间存在交联种。我们的结果表明，OCP的n端结构域与两个异藻蓝蛋白三聚体在藻胆小体核心基柱上形成的位点密切相关。本报告有助于了解在蓝藻非光化学猝灭过程中OCP的活化机制和OCP的结构结合位点。
The Orange Carotenoid Protein (OCP) plays a similar photoprotective role in cyanobacterial photosynthesis to that of non-photochemical quenching in higher plants. Under high-light conditions, OCP binds to the phycobilisome (PBS) and reduces energy transfer to the photosystems. The protective cycle starts from a light-induced activation of OCP. Detailed information on the molecular mechanism of this process as well as the subsequent recruitment of active OCP to the phycobilisome is not known. We report here our investigation on OCP photoactivation from the cyanobacterium Synechocystis sp. PCC 6803 by using a combination of native mass spectrometry (MS) and protein cross-linking. We demonstrate that Native MS is able to capture OCP with its intact pigment and further reveal that OCP undergoes a dimer-tomonomer transition upon light illumination. The reversion of activated form of OCP to inactive, dark form was also observed by using native MS. Furthermore, in vitro reconstitution of OCP and PBS allowed to perform protein chemical cross-linking experiments. LC-MS/MS analysis identified cross-linking species between OCP and the PBS core components. Our result indicates that the N-terminal domain of OCP is closely involved in the association with a site formed by two allophycocyanin trimers in the basal cylinders of the phycobilisome core. This report helps to understand the activation mechanism of OCP and the structural binding site of OCP during the cyanobacterial non-photochemical quenching process.
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DOI：
10.1016/j.jphotobiol.2013.05.014
发表时间：
2013-08-05
影响因子：
5.4
作者：
Stadnichuk, Igor N.;Yanyushin, Mikhail F.;Paschenko, Vladimir Z.
通讯作者：
Paschenko, Vladimir Z.
影响因子：
15
作者：
Tian, Lijin;van Stokkum, Ivo H. M.;van Amerongen, Herbert
通讯作者：
van Amerongen, Herbert
影响因子：
3.1
作者：
Kirilovsky, Diana;Kerfeld, Cheryl A.
通讯作者：
Kerfeld, Cheryl A.
影响因子：
14.8
作者：
通讯作者：
--
DOI：
10.1126/science.1242321
发表时间：
2013-11-29
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Liu H;Zhang H;Niedzwiedzki DM;Prado M;He G;Gross ML;Blankenship RE
通讯作者：
Blankenship RE