Sekiguchi Hiroshi, Arakawa Hideo, Taguchi Hideki, Ito Takeshi, Kokawa Ryohei, Ikai Atsushi
Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, Midori-ku, Yokohama, Kanagawa 226-8501, Japan.
Biophys J. 2003 Jul;85(1):484-90. doi: 10.1016/S0006-3495(03)74493-2.
We investigated the interaction between GroEL and a denatured protein from a mechanical point of view using an atomic force microscope. Pepsin was bound to an atomic force microscope probe and used at a neutral pH as an example of denatured proteins. To measure a specific and delicate interaction force, we obtained force curves without pressing the probe onto GroEL molecules spread on a mica surface. Approximately 40 pN of tensile force was observed for approximately 10 nm while pepsin was pulled away from the chaperonin after a brief contact. This length of force duration corresponding to the circumference of GroEL's interior cavity was shortened by the addition of ATP. The relation between the observed mechanical parameters and the chaperonin's refolding function is discussed.
我们使用原子力显微镜从力学角度研究了GroEL与变性蛋白之间的相互作用。胃蛋白酶被固定在原子力显微镜探针上，并在中性pH条件下用作变性蛋白的示例。为了测量特定且微妙的相互作用力，我们在不将探针压在铺展在云母表面的GroEL分子上的情况下获得了力曲线。在短暂接触后，当胃蛋白酶从伴侣蛋白上被拉开时，在大约10纳米的距离内观察到了约40皮牛的拉力。对应于GroEL内部腔室周长的这种力持续时间长度因添加ATP而缩短。本文讨论了观察到的力学参数与伴侣蛋白的重折叠功能之间的关系。
