Fast Photochemical Oxidation of Proteins Maps the Topology of Intrinsic Membrane Proteins: Light-Harvesting Complex 2 in a Nanodisc.
Fast Photochemical Oxidation of Proteins Maps the Topology of Intrinsic Membrane Proteins: Light-Harvesting Complex 2 in a Nanodisc.
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DOI：
10.1021/acs.analchem.6b01945
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发表时间：
2016-09-06
影响因子：
7.4
通讯作者：
Gross, Michael L.
中科院分区：
文献类型：
--
作者：
Lu, Yue;Zhang, Hao;Niedzwiedzki, Dariusz M.;Jiang, Jing;Blankenship, Robert E.;Gross, Michael L.
尽管膜蛋白是光合作用和其他生物过程的关键参与者，但许多膜蛋白缺乏高分辨率结构。在获得高分辨率结构之前，我们正在研究MS-based足迹是否可以通过跟踪配体结合、pH变化和膜结合时发生的结构变化来提供粗粒度的蛋白质结构。我们的平台通过结合MS-based足迹，特别是蛋白质的快速光化学氧化（FPOP）和脂质纳米盘来探测膜蛋白的拓扑和构象，脂质纳米盘比广泛使用的洗涤剂胶束更类似于天然膜环境。我们在这里描述的结果表明，蛋白质的外膜区域被OH自由基更重的足迹，而跨越脂质双分子层的区域对标记保持惰性。与洗涤剂胶束相比，纳米圆盘通常表现出对膜蛋白的更多保护，而对存在于膜外的蛋白质残基的屏蔽作用较小。将蛋白质固定在纳米圆盘中并结合FPOP方法进行足迹标记是一种可行的方法，可以绘制膜外蛋白表面，甚至在氨基酸水平上，并阐明膜蛋白的内在拓扑结构。
Although membrane proteins are crucial participants in photosynthesis and other biological processes, many lack high-resolution structures. Prior to achieving a high-resolution structure, we are investigating whether MS-based footprinting can provide coarse-grained protein structure by following structural changes that occur upon ligand binding, pH change, and membrane binding. Our platform probes topology and conformation of membrane proteins by combining MS-based footprinting, specifically fast photochemical oxidation of proteins (FPOP), and lipid Nanodiscs, which more similar to the native membrane environment than are the widely used detergent micelles. We describe here results that show a protein’s outer membrane regions are more heavily footprinted by OH radicals whereas the regions spanning the lipid bilayer remain inert to the labeling. Nanodiscs generally exhibit more protection of membrane proteins compared to detergent micelles and less shielding to those protein residues that exist outside the membrane. The combination of immobilizing the protein in Nanodiscs and footprinting with the FPOP approach is a feasible approach to map extra-membrane protein surfaces, even at the amino-acid level, and to illuminate intrinsic membrane protein topology.
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影响因子：
4.8
作者：
Bayburt, Timothy H.;Vishnivetskiy, Sergey A.;Gurevich, Vsevolod V.
通讯作者：
Gurevich, Vsevolod V.
影响因子：
3.9
作者：
Bond, PJ;Cuthbertson, J;Sansom, MSP
通讯作者：
Sansom, MSP
影响因子：
5.8
作者：
Hennerdal, Aron;Elofsson, Arne
通讯作者：
Elofsson, Arne
DOI：
10.1073/pnas.78.9.5583
发表时间：
1981-01-01
期刊：
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA-BIOLOGICAL SCIENCES
影响因子：
--
作者：
CLAYTON, RK;CLAYTON, BJ
通讯作者：
CLAYTON, BJ
影响因子：
14.9
作者：
UniProt Consortium
通讯作者：
UniProt Consortium