title: Analysis of the aplyronine A-induced protein-protein interaction between actin and tubulin by surface plasmon resonance.
abstract: The antitumor macrolide aplyronine A induces protein-protein interaction (PPI) between actin and tubulin to exert highly potent biological activities. The interactions and binding kinetics of these molecules were analyzed by the surface plasmon resonance with biotinylated aplyronines or tubulin as ligands. Strong binding was observed for tubulin and actin with immobilized aplyronine A. These PPIs were almost completely inhibited by one equivalent of either aplyronine A or C, or mycalolide B. In contrast, a non-competitive actin-depolymerizing agent, latrunculin A, highly accelerated their association. Significant binding was also observed for immobilized tubulin with an actin-aplyronine A complex, and the dissociation constant KD was 1.84μM. Our method could be used for the quantitative analysis of the PPIs between two polymerizing proteins stabilized with small agents.
authors.affiliation: Graduate School of Pure and Applied Sciences, University of Tsukuba, 1-1-1 Tennodai, Tsukuba 305-8571, Japan.
authors.forename: Yuichiro
authors.initials: Y
authors.lastname: Hirayama
authors.affiliation: Graduate School of Pure and Applied Sciences, University of Tsukuba, 1-1-1 Tennodai, Tsukuba 305-8571, Japan.
authors.forename: Kota
authors.initials: K
authors.lastname: Yamagishi
authors.affiliation: Research Institute of Green Science and Technology, Shizuoka University, 836 Ohya, Suruga, Shizuoka 422-8529, Japan.
authors.forename: Tomohiro
authors.initials: T
authors.lastname: Suzuki
authors.affiliation: Research Institute of Green Science and Technology, Shizuoka University, 836 Ohya, Suruga, Shizuoka 422-8529, Japan; Graduate School of Agriculture, Shizuoka University, 836 Ohya, Suruga, Shizuoka 422-8529, Japan; Graduate School of Science and Technology, Shizuoka University, 836 Ohya, Suruga, Shizuoka 422-8529, Japan.
authors.forename: Hirokazu
authors.initials: H
authors.lastname: Kawagishi
authors.affiliation: Graduate School of Pure and Applied Sciences, University of Tsukuba, 1-1-1 Tennodai, Tsukuba 305-8571, Japan; Tsukuba Research Center for Interdisciplinary Materials Science (TIMS), University of Tsukuba, 1-1-1 Tennodai, Tsukuba 305-8571, Japan; JST-PRESTO, 1-1-1 Tennodai, Tsukuba 305-8571, Japan. Electronic address: mkita@chem.tsukuba.ac.jp.
authors.forename: Masaki
authors.initials: M
authors.lastname: Kita
authors.affiliation: Graduate School of Pure and Applied Sciences, University of Tsukuba, 1-1-1 Tennodai, Tsukuba 305-8571, Japan. Electronic address: kigoshi@chem.tsukuba.ac.jp.
authors.forename: Hideo
authors.initials: H
authors.lastname: Kigoshi
mesh_terms: D000199:Actins / Q000378:metabolism*; D000818:Animals; D000970:Antineoplastic Agents / Q000302:isolation & purification / Q000494:pharmacology*; D001048:Aplysia / Q000737:chemistry; D006367:HeLa Cells; D006801:Humans; D018942:Macrolides / Q000302:isolation & purification / Q000494:pharmacology*; D060066:Protein Interaction Maps / Q000187:drug effects*; D020349:Surface Plasmon Resonance; D014404:Tubulin / Q000378:metabolism*
chemical_list: D000199:Actins; D000970:Antineoplastic Agents; D018942:Macrolides; D014404:Tubulin; C104057:aplyronine A
keywords: Actin; Antitumor natural products; Protein–protein interaction; Surface plasmon resonance; Tubulin