TY - JOUR
T1 - Electron microscopic visualization of the filament binding mode of actin-binding proteins
AU - Ito, Takuto
AU - Hirayama, Tasuku
AU - Taki, Masayasu
AU - Iyoshi, Shohei
AU - Dai, Shuheng
AU - Takeda, Shuichi
AU - Kimura-Sakiyama, Chieko
AU - Oda, Toshiro
AU - Yamamoto, Yukio
AU - Maéda, Yuichiro
AU - Narita, Akihiro
N1 - Funding Information:
This work was supported by The Uehara Memorial Foundation , Takeda Science Foundation and by a Grant-in-Aid for Scientific Research (S:20227008) from The Ministry of Education, Culture, Sports, Science and Technology of the Japanese Government . We thank Prof. Masayuki Kajitani (Teikyo Univ.) for providing the tobacco mosaic virus, Prof. Masao Miki (Fukui Univ.) for providing tropomyosin, Dr. Yoshiyuki Matsuura (Nagoya Univ.) for providing an expression vector of profilin and Dr. Kayo Maeda (Nagoya Univ.) for providing cDNAs of troponin subunits.
PY - 2011/4/22
Y1 - 2011/4/22
N2 - 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.
AB - 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.
KW - actin filament
KW - end-binding proteins
KW - gold nanoparticle
KW - image analysis
KW - transmission electron microscopy
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U2 - 10.1016/j.jmb.2011.01.054
DO - 10.1016/j.jmb.2011.01.054
M3 - Article
C2 - 21315081
AN - SCOPUS:79953243387
SN - 0022-2836
VL - 408
SP - 26
EP - 39
JO - Journal of Molecular Biology
JF - Journal of Molecular Biology
IS - 1
ER -