Electron Microscopic Visualization of the Filament Binding Mode of Actin-Binding Proteins

被引:3
作者
Ito, Takuto [1 ,2 ,3 ]
Hirayama, Tasuku [4 ]
Taki, Masayasu [4 ,5 ]
Iyoshi, Shohei [4 ]
Dai, Shuheng [1 ,2 ,3 ]
Takeda, Shuichi [1 ,2 ,3 ]
Kimura-Sakiyama, Chieko [1 ,2 ,3 ]
Oda, Toshiro [3 ,6 ]
Yamamoto, Yukio [5 ]
Maeda, Yuichiro [1 ,2 ,3 ]
Narita, Akihiro [1 ,2 ,3 ]
机构
[1] Nagoya Univ, Struct Biol Res Ctr, Grad Sch Sci, Chikusa Ku, Nagoya, Aichi 4648601, Japan
[2] Nagoya Univ, Div Biol Sci, Grad Sch Sci, Chikusa Ku, Nagoya, Aichi 4648601, Japan
[3] Japan Sci & Technol Agcy, ERATO Actin Filament Dynam Project, Mikazuki, Hyogo 6795148, Japan
[4] Kyoto Univ, Grad Sch Human & Environm Studies, Sakyo Ku, Kyoto 6068501, Japan
[5] Kyoto Univ, Grad Sch Global Environm Studies, Sakyo Ku, Kyoto 6068501, Japan
[6] RIKEN SPring 8 Ctr, RIKEN Harima Inst, Xray Struct Anal Team, Mikazuki, Hyogo 6795148, Japan
关键词
actin filament; end-binding proteins; transmission electron microscopy; gold nanoparticle; image analysis; UNDECAGOLD CLUSTERS; SKELETAL-MUSCLE; CAPPING PROTEIN; TAGGED PROTEINS; GOLD CLUSTERS; BARBED-END; NUCLEATION; TROPOMYOSIN; COMPLEXES; PROFILIN;
D O I
10.1016/j.jmb.2011.01.054
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
A large number of actin-binding proteins (ABPs) regulate various kinds of cellular events in which the superstructure of the actin cytoskeleton is dynamically changed. Thus, to understand the actin dynamics in the cell, the mechanisms of actin regulation by ABPs must be elucidated. Moreover, it is particularly important to identify the side, barbed-end or pointed-end ABP binding sites on the actin filament. However, a simple, reliable method to determine the ABP binding sites on the actin filament is missing. Here, a novel electron microscopic method for determining the ABP binding sites is presented. This approach uses a gold nanoparticle that recognizes a histidine tag on an ABP and an image analysis procedure that can determine the polarity of the actin filament. This method will facilitate future study of ABPs. (C) 2011 Published by Elsevier Ltd.
引用
收藏
页码:26 / 39
页数:14
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