Super Complexes and Trans-Membrane Signaling of the Cytochrome b6f Complex

细胞色素 b6f 复合物的超级复合物和跨膜信号传导

• 批准号: 9291998
• 负责人: William A. Cramer
• 金额: $ 15.21万
• 依托单位: PURDUE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1987
• 资助国家: 美国
• 起止时间: 1987-04-01 至 2017-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9291998
• 关键词: Achievement; Active Biological Transport; Adrenergic Receptor; Affect; Affinity; Binding; Binding Sites; Biological Assay; Carrier Proteins; Charge; Chlorophyll; Circular Dichroism; Complex; Coupled; Crystallization; Cytochrome a; Cytochrome bc1 Complex; Cytochromes; Cytochromes b; Data; Databases; Dependence; Detergents; Deuterium; Disease; Drug Targeting; Electron Transport; Electronic Mail; Environment; G-Protein-Coupled Receptors; Goals; Green Algae; Health; Heme; Heterogeneity; Higher Order Chromatin Structure; Hydrogen; Letters; Lipid Binding; Lipids; Lipoproteins; Liposomes; Mass Spectrum Analysis; Mediating; Membrane; Membrane Proteins; Methods; Mitochondria; Nostoc; Occupations; Oxidation-Reduction; Pathway interactions; Phosphotransferases; Photosystem I; Preparation; Property; Protein Kinase; Proteins; Protons; Quinones; Reaction; Resolution; Respiratory Chain; Role; Side; Signal Transduction; Site; Source; Staging; Structure; System; Testing; Water; base; electric field; electrical potential; electron donor; experience; improved; interest; monomer; oxidation; protein complex; protein function; protein structure; reconstitution; response; signal processing; stoichiometry

DESCRIPTION (provided by applicant): (1) The system. The cytochrome b6f complex is one of only 60 independent hetero-oligomeric membrane proteins in the data bank. (2) Recent achievements, structure-function: (a) a newly recognized unique heme in the complex was characterized; (b) the quinone dependent pathway for proton translocation was described at a resolution of 2.7 A (c) a unique function of the bound chlorophyll in occluding the quinone entry portal was demonstrated; (d) detailed lipid analysis was undertaken, conservation shown of many lipid functions between the b6f complex and the mitochondrial cytochrome bc1 complex, and (e) conformational changes shown to be induced by anionic lipid. (3) Proposed studies: (a) Using His-tagged cytochrome b6f complex derived from C. reinhardtii, (b) crystal structures of higher order ("super")-complexes of the electron transport complexes in the photosynthetic electron transport chain will be sought, following the precedent set for such super-complexes in the respiratory chain. A starting point is utilization of an isolated complex of the 230 kDa cytochrome b6f complex with the 1 MDa trimeric photosystem I reaction center. Elucidation of the structure of such a complex would likely describe new pathways for electronic communication between the complexes via quinones and soluble electron transport proteins, and be used to define the interactions and functions of the lipids internal to the cytochrome complex. (c) The stoichiometry and arrangement of the lipids, and their effects on the dynamics of the cytochrome complex, will be analyzed by native and hydrogen-deuterium mass spectroscopy. (d) Assembly; role of lipids. To test the role of the lipids in separating different domains of the cytochrome complex, mass spectroscopic analysis, particularly native MS, will be applied to analysis of lipid interactions at different stages of assembly of the b6f complex. (e Dielectric heterogeneity of the b6f complex; dependence of structure on electric field. Our studies imply that the internal dielectric constant within the b6f complex is heterogeneous, i. e., different between the different heme pairs within the complex. Virtually all membrane proteins, certainly those involved in energy-transduction and active transport, function in the presence of large electrical fields. With the long-term goal of determining the effect of electric fields on membrane protein structures, the effect of such fields on cytochrome spectra, redox properties, and secondary structure assayed by far-UV circular dichroism spectra, will be studied. (f) Trans- membrane signaling; structure of the signaling complex. The redox signaling system in the cytochrome b6f complex uses a unique trans-membrane signaling mechanism dependent upon oxidation of plastoquinol on the electro-positive side of the complex. A crystal structure will be sought of the trans-membrane signaling complex consisting of the b6f complex reconstituted with the protein kinase. These studies are directed not only to better crystallographic resolution and understanding of the structure of the cytochrome complex as a hetero- oligomeric membrane lipoprotein that is relevant to its function of trans-membrane signaling.

描述（申请人提供）：（1）系统。细胞色素b6 f复合物是数据库中仅有的60种独立的异源寡聚膜蛋白之一。(2)结构-功能方面的最新进展：（a）确定了复合物中一个新发现的独特血红素，（B）描述了醌依赖的质子转运途径，分辨率为2.7 A，（c）证明了结合叶绿素在封闭醌进入通道中的独特功能;（d）进行了详细的脂质分析，显示b6 f复合物和线粒体细胞色素bc 1复合物之间许多脂质功能的保守性，和（e）显示由阴离子脂质诱导的构象变化。(3)建议的研究：（a）使用来自C. reinhardtii，（B）将遵循呼吸链中电子传递复合物的高级（“超级”）复合物的先例，寻找光合作用电子传递链中电子传递复合物的高级（“超级”）复合物的晶体结构。起点是利用230 kDa细胞色素b6 f复合物与1 MDa三聚体光系统I反应中心的分离复合物。阐明这种复合物的结构可能会描述新的途径，通过醌类和可溶性电子传递蛋白的复合物之间的电子通信，并被用来定义的相互作用和功能的脂质内部的细胞色素复合物。(c)脂质的化学计量和排列，以及它们对细胞色素复合物动力学的影响，将通过天然和氢-氘质谱法进行分析。(d)组装;脂质的作用。为了测试脂质在分离细胞色素复合物的不同结构域中的作用，质谱分析，特别是天然MS，将应用于分析b6 f复合物组装的不同阶段的脂质相互作用。（e）b6 f复合物的介电不均匀性;结构对电场的依赖性。我们的研究表明，b6 f复合物的内部介电常数是不均匀的，即。例如， 不同的血红素对之间的差异。事实上，所有的膜蛋白，当然是那些参与能量转导和主动运输的蛋白，在大电场的存在下发挥作用。随着确定电场对膜蛋白结构的影响的长期目标，将研究这些领域对细胞色素光谱，氧化还原性质和二级结构测定的远紫外圆二色光谱的影响。(f)跨膜信号传导;信号复合物的结构。细胞色素b6 f复合物中的氧化还原信号传导系统使用依赖于复合物的正电侧上的质体醌醇的氧化的独特的跨膜信号传导机制。将寻求由用蛋白激酶重构的b6 f复合物组成的跨膜信号传导复合物的晶体结构。这些研究不仅涉及更好的晶体学分辨率和理解作为异源寡聚膜脂蛋白的细胞色素复合物的结构，其与其跨膜信号传导功能相关。

项目成果

Thylakoid membrane protein topography: transmembrane orientation of the chloroplast cytochrome b-559 psbE gene product.

类囊体膜蛋白形貌：叶绿体细胞色素 b-559 psbE 基因产物的跨膜方向。

• DOI: 10.1021/bi00426a002
• 发表时间: 1988
• 期刊: Biochemistry
• 影响因子: 2.9
• 作者: Tae,GS;Black,MT;Cramer,WA;Vallon,O;Bogorad,L
• 通讯作者: Bogorad,L

Electrostatic destabilization of the cytochrome b6f complex in the thylakoid membrane.

类囊体膜中细胞色素 b6f 复合物的静电不稳定。

• DOI: 10.1002/j.1460-2075.1991.tb07824.x
• 发表时间: 1991
• 期刊: The EMBO journal
• 作者: Szczepaniak,A;Huang,D;Keenan,TW;Cramer,WA
• 通讯作者: Cramer,WA

Note: Small anaerobic chamber for optical spectroscopy.

注意：用于光谱学的小型厌氧室。

• DOI: 10.1063/1.4932183
• 发表时间: 2015
• 期刊: The Review of scientific instruments
• 作者: Chauvet,AdrienAP;Agarwal,Rachna;Cramer,WilliamA;Chergui,Majed
• 通讯作者: Chergui,Majed

Topography of the chloroplast cytochrome b6: orientation of the cytochrome and accessibility of the lumen-side interhelix loops.

叶绿体细胞色素 b6 的形貌：细胞色素的方向和管腔侧螺旋间环的可及性。

• DOI: 10.1515/znc-1989-5-619
• 发表时间: 1989
• 期刊: Zeitschrift fur Naturforschung. C, Journal of biosciences
• 作者: Szczepaniak,A;Black,MT;Cramer,WA
• 通讯作者: Cramer,WA

Electrostatic effects on electron-transfer kinetics in the cytochrome f-plastocyanin complex.

静电对细胞色素 f-质体蓝蛋白复合物中电子转移动力学的影响。

• DOI: 10.1016/s0006-3495(97)78351-6
• 发表时间: 1997
• 期刊: Biophysical journal
• 影响因子: 3.4
• 作者: Soriano,GM;Cramer,WA;Krishtalik,LI
• 通讯作者: Krishtalik,LI