title: Electron microscopic visualization of the filament binding mode of actin-binding proteins.
abstract: 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.
authors.affiliation: Structural Biology Research Center and Division of Biological Sciences, Graduate School of Science, Nagoya University, Furo-cho, Chikusa-ku, Nagoya 464-8601, Japan.
authors.forename: Takuto
authors.initials: T
authors.lastname: Ito
authors.forename: Tasuku
authors.initials: T
authors.lastname: Hirayama
authors.forename: Masayasu
authors.initials: M
authors.lastname: Taki
authors.forename: Shohei
authors.initials: S
authors.lastname: Iyoshi
authors.forename: Shuheng
authors.initials: S
authors.lastname: Dai
authors.forename: Shuichi
authors.initials: S
authors.lastname: Takeda
authors.forename: Chieko
authors.initials: C
authors.lastname: Kimura-Sakiyama
authors.forename: Toshiro
authors.initials: T
authors.lastname: Oda
authors.forename: Yukio
authors.initials: Y
authors.lastname: Yamamoto
authors.forename: Yuichiro
authors.initials: Y
authors.lastname: Maéda
authors.forename: Akihiro
authors.initials: A
authors.lastname: Narita
mesh_terms: D000199:Actins / Q000737:chemistry / Q000378:metabolism*; D001665:Binding Sites; D003599:Cytoskeleton / Q000737:chemistry / Q000378:metabolism; D017612:Gold Compounds / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D053768:Metal Nanoparticles / Q000737:chemistry; D008840:Microfilament Proteins / Q000737:chemistry / Q000378:metabolism* / Q000648:ultrastructure; D008854:Microscopy, Electron; D011485:Protein Binding
chemical_list: D000199:Actins; D017612:Gold Compounds; D008840:Microfilament Proteins