2. Academic Validation
  3. Multimodal biopanning of T7 phage-displayed peptides reveals angiomotin as a potential receptor of the anti-angiogenic macrolide Roxithromycin

Multimodal biopanning of T7 phage-displayed peptides reveals angiomotin as a potential receptor of the anti-angiogenic macrolide Roxithromycin

  • Eur J Med Chem. 2015 Jan 27:90:809-21. doi: 10.1016/j.ejmech.2014.12.015.
Kaori Takakusagi 1,  Yoichi Takakusagi 2,  Takahiro Suzuki 3,  Aya Toizaki 3,  Aiko Suzuki 1,  Yaichi Kawakatsu 1,  Madoka Watanabe 1,  Yukihiro Saito 4,  Ryushi Fukuda 4,  Atsuo Nakazaki 3,  Susumu Kobayashi 3,  Kengo Sakaguchi 1,  Fumio Sugawara 5
Affiliations

Affiliations

  • 1 Department of Applied Biological Science, Faculty of Science and Technology, Tokyo University of Science, 2641 Yamazaki, Noda, Chiba 278-8510, Japan.
  • 2 Department of Applied Biological Science, Faculty of Science and Technology, Tokyo University of Science, 2641 Yamazaki, Noda, Chiba 278-8510, Japan. Electronic address: tkksg@rs.noda.tus.ac.jp.
  • 3 Faculty of Pharmaceutical Sciences, Tokyo University of Science, 2641 Yamazaki, Noda, Chiba 278-8510, Japan.
  • 4 Meiwafosis Co. Ltd., 1-14-2 Shinjuku, Tokyo 160-0022, Japan.
  • 5 Department of Applied Biological Science, Faculty of Science and Technology, Tokyo University of Science, 2641 Yamazaki, Noda, Chiba 278-8510, Japan. Electronic address: sugawara@rs.noda.tus.ac.jp.
PMID: 25528335 DOI: 10.1016/j.ejmech.2014.12.015
Abstract

Roxithromycin (RXM) is a semi-synthetic fourteen-membered macrolide Antibiotic that shows anti-angiogenic activity in solid tumors. In the present study, we conducted biopanning of T7 phage-displayed Peptides either on a 96-well formatted microplate, a flow injection-type quartz-crystal microbalance (QCM) biosensor, or a cuvette-type QCM. RXM-selected Peptides of different sequence, length and number were obtained from each mode of screening. Subsequent bioinformatics analysis of the RXM-selected Peptides consistently gave positive scores for the extracellular domain (E458-T596) of angiomotin (Amot), indicating that this may comprise a binding region for RXM. Bead pull down assay and QCM analysis confirmed that RXM directly interacts with Amot via the screen-guided region, which also corresponds to the binding site for the endogenous anti-angiogenic inhibitor angiostatin (Anst). Thus, multimodal biopanning of T7PD revealed that RXM binds to the extracellular domain on Amot as a common binding site with Anst, leading to inhibition of angiogenesis-dependent tumor growth and metastasis. These data might explain the molecular basis underlying the mechanism of action for the anti-angiogenic activity of RXM.

Keywords

Angiogenesis; Angiomotin; QCM-D; RELIC; Roxithromycin; T7 phage display.

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