2. Academic Validation
  3. Analysis of the aplyronine A-induced protein-protein interaction between actin and tubulin by surface plasmon resonance

Analysis of the aplyronine A-induced protein-protein interaction between actin and tubulin by surface plasmon resonance

  • Bioorg Med Chem. 2016 Jun 15;24(12):2809-14. doi: 10.1016/j.bmc.2016.04.049.
Yuichiro Hirayama 1,  Kota Yamagishi 1,  Tomohiro Suzuki 2,  Hirokazu Kawagishi 3,  Masaki Kita 4,  Hideo Kigoshi 5
Affiliations

Affiliations

  • 1 Graduate School of Pure and Applied Sciences, University of Tsukuba, 1-1-1 Tennodai, Tsukuba 305-8571, Japan.
  • 2 Research Institute of Green Science and Technology, Shizuoka University, 836 Ohya, Suruga, Shizuoka 422-8529, Japan.
  • 3 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.
  • 4 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.
  • 5 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.
PMID: 27161875 DOI: 10.1016/j.bmc.2016.04.049
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.

Keywords

Actin; Antitumor natural products; Protein–protein interaction; Surface plasmon resonance; Tubulin.

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