NMR STUDIES OF BIOLOGICAL MEMBRANES

生物膜的核磁共振研究

• 批准号: 6519223
• 负责人: JUDITH HERZFELD
• 金额: $ 25.87万
• 依托单位: BRANDEIS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1985
• 资助国家: 美国
• 起止时间: 1985-09-30 至 2004-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6519223
• 关键词: Halobacteriaceae; Schiff bases; active sites; active transport; bacteriorhodopsins; bioimaging /biomedical imaging; cell membrane; chromophore; conformation; electronic spectra; hydrogen transport; nuclear magnetic resonance spectroscopy; photochemistry; protein folding; protein structure function; water

Description: (Verbatim from the applicant's abstract) A distinctive feature of living cells is the facility with which they convert energy from one form to another. Central among these energy transactions is the active transport of ions across cell membranes. In recent years, significant progress has been made in elucidating the structures of various ion pumps. However, the molecular mechanisms by which vectorial ion motion is enforced remain unknown. Our long-term goal is to understand how proteins carry out this essential aspect of energy transduction. A particularly propitious system for this purpose is the light-driven proton pump, bacteriorhodopsin. Key steps in the photocycle have been identified and the structure of the resting state has been established by diffraction methods (with the exception of a few disordered, packing sensitive, or hydration sensitive regions). Our goal is to obtain a detailed picture of the active site of the molecule as it evolves through the critical steps of the photocycle. In particular, we are concerned with the structural changes that occur around the Schiff base of the retinal chromophore while it is deprotonated (i.e., during the M stage of the photocycle), because these changes prevent the proton that is released to the extracellular side from returning when the Schiff base reprotonates. Thus we are looking specifically for a switch in the connectivity of the deprotonated Schiff base between transport pathways on the two sides of the membrane and clues to the source of the irreversibility of this switch. Since our work was last funded, we have identified early and late M intermediates and learned how to trap them at levels suitable for solid state NMR studies. In addition, the last few years have seen a dramatic expansion of the range and power of solid state NMR techniques and we have developed procedures for new variations in the isotopic labeling of bacteriorhodopsin that will allow us to take advantage of the new spectroscopy. Bringing these elements together, we will characterize the spatial relationships and chemical exchange connectivities that define the active site and how they change in the critical photocycle intermediates.

描述：（逐字摘自申请人的摘要） 活细胞是将能量从一种形式转化为一种形式的设施 其他。这些能源交易的核心是主动运输 离子穿过细胞膜。近年来取得了重大进展 阐明各种离子泵的结构。然而，分子 强制矢量离子运动的机制仍然未知。我们的 长期目标是了解蛋白质如何执行这一重要方面 能量转换。 用于此目的的一个特别有利的系统是光驱动质子 泵，细菌视紫红质。光循环中的关键步骤已被确定并 通过衍射方法建立了静止态的结构 （除少数无序、包装敏感或水合 敏感区域）。我们的目标是获得活动站点的详细图片 分子在光循环的关键步骤中演化的过程。在 特别是，我们关注周围发生的结构性变化 视网膜发色团去质子化时（即在 光循环的 M 阶段），因为这些变化阻止了质子 当希夫碱返回时被释放到细胞外侧 重质子化。因此，我们正在专门寻找连接中的开关 两侧传输路径之间的去质子化席夫碱 膜和此开关不可逆性来源的线索。 自从我们的工作上次获得资助以来，我们已经确定了早期和晚期 M 中间体并学习如何将它们捕获在适合固态的水平 核磁共振研究。此外，在过去几年中， 固态核磁共振技术的范围和能力，我们已经开发 细菌视紫红质同位素标记新变化的程序 这将使我们能够利用新的光谱学。带这些 元素在一起，我们将表征空间关系和化学 定义活动站点的交换连接以及它们如何在 关键的光循环中间体。

项目成果

Determination of membrane protein structure by rotational resonance NMR: bacteriorhodopsin.

通过旋转共振核磁共振测定膜蛋白结构：细菌视紫红质。

• 发表时间: 1991-02-15
• 作者: Creuzet, F;McDermott, A;Gebhard, R;van der Hoef, K;Spijker;Herzfeld, J;Lugtenburg, J;Levitt, M H;Griffin, R G
• 通讯作者: Griffin, R G

Nuclear magnetic resonance study of the Schiff base in bacteriorhodopsin: counterion effects on the 15N shift anisotropy.

细菌视紫红质中希夫碱的核磁共振研究：抗衡离子对 15N 位移各向异性的影响。

• 发表时间: 1989-04-18
• 影响因子: 2.9
• 作者: de Groot, H J;Harbison, G S;Herzfeld, J;Griffin, R G
• 通讯作者: Griffin, R G

Magnetic birefringence studies of dilute purple membrane suspensions.

稀紫色膜悬浮液的磁双折射研究。

• 发表时间: 1985-02
• 影响因子: 3.4
• 作者: Lewis, B A;Rosenblatt, C;Griffin, R G;Courtemanche, J;Herzfeld, J
• 通讯作者: Herzfeld, J

Solid-state 13C NMR study of tyrosine protonation in dark-adapted bacteriorhodopsin.

暗适应细菌视紫红质中酪氨酸质子化的固态 13C NMR 研究。

• DOI: 10.1021/bi00475a022
• 发表时间: 1990-06-12
• 期刊: Biochemistry
• 影响因子: 2.9
• 作者: J. Herzfeld;S. Das Gupta;M. Farrar;G. Harbison;A. McDermott;S. Pelletier;D. Raleigh;S. O. Smith;C. Winkel;J. Lugtenburg
• 通讯作者: J. Lugtenburg

Solid-state 13C NMR detection of a perturbed 6-s-trans chromophore in bacteriorhodopsin.

细菌视紫红质中受干扰的 6-s-反式发色团的固态 13C NMR 检测。

• DOI: 10.1021/bi00345a031
• 发表时间: 1985-11-19
• 期刊: Biochemistry
• 影响因子: 2.9
• 作者: G. Harbison;Steven O. Smith;J. A. Pardoen;J. Courtin;J. Lugtenburg;J. Herzfeld;R. Mathies;R. G. Griffin
• 通讯作者: R. G. Griffin