Pulling and pushing protein molecules by AFM
Pulling and pushing protein molecules by AFM
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DOI：
10.2174/157341307779940535
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发表时间：
2007-02-01
影响因子：
1.5
通讯作者：
Sekiguchi, Hiroshi
中科院分区：
文献类型：
--
作者：
Ikai, Atsushi;Afrin, Rehana;Sekiguchi, Hiroshi
本文综述了近年来在分子水平上对表面固定化蛋白质和包埋在脂质双层中的膜结合蛋白质刚性的研究进展。一些细节和重点给出了在我们的实验室在过去几年中所做的工作。蛋白质分子通过共价交联剂固定在固体表面上，另一侧固定在原子力显微镜的探针上，使我们能够将单个蛋白质分子拉或推到指定的方向。在拉的时候，蛋白质分子的内部结构被机械地打开，在推的时候，它被压缩变形，直到它被压平。这些实验以两种不同的方式揭示了蛋白质分子折叠结构的机械刚性。在配体-蛋白质相互作用领域，讨论了新近引入的无压缩方法的优点和存在的问题。当蛋白质分子嵌入脂质膜中时，通过将其从膜中拉出可以获得关于其对膜的锚定力的信息。实验已经在固体表面上形成的脂质双层或活细胞表面上进行。膜蛋白的提取过程中往往伴随着一条尾随在靶蛋白后面的薄脂质系链的挤压拉力。将给出从红细胞膜表面的系链形成过程的简短回顾。
This paper reviews a recent progress of molecular level studies on the rigidity of surface immobilized as well as membrane bound proteins embedded in the lipid bilayer. Some details and emphasis are given to the work done in our laboratory in the last few years. Immobilization of protein molecules on a solid surface through covalent crosslinkers on one side and to the probe of the atomic force microscope on the other enabled us to pull or push a single protein molecule to specified directions. On pulling, the internal structure of the protein molecule is mechanically opened up and, on pushing, it is compressively deformed until it is flattened out. Such experiments reveal the mechanical rigidity of the folded structure of a protein molecule in two different ways. In the field of ligand-protein interaction, some merits and problems of newly introduced compression free method are discussed. When a protein molecule is embedded in a lipid membrane, information on its anchoring force to the membrane can be obtained by pulling it out from the membrane. Experiments have been done either on a lipid bilayer formed on a solid surface, or on the surface of live cells. The extraction process of membrane proteins is often accompanied by extrusion of a thin lipid tether trailing behind the target protein of the tensile force. A short review of the tether forming process from the red cell membrane surface will be given.