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Recombinant pigment epithelium-derived factor PEDF binds vascular endothelial growth factor receptors 1 and 2

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Abstract

Angiogenesis, or the formation of new blood vessels, is stimulated by angiogenic factors such as vascular endothelial growth factor (VEGF). Pigment epithelium-derived factor (PEDF) is a potent inhibitor of angiogenesis. To explore the mechanism by which PEDF acts, recombinant PEDF was expressed with a 6x-His tag (for purification) and a green fluorescent protein (GFP) tag. The PEDF fusion protein was confirmed to be active in inhibition of endothelial cell proliferation and migration. Direct binding of PEDF to both vascular endothelial growth factor receptor-1 (VEGFR-1) and VEGFR-2 was demonstrated in an in vitro assay similar to an enzyme-linked immunosorbent assay (ELISA). PEDF was shown by immune-confocal microscopy to be localized within treated endothelial cells. When VEGF-stimulated endothelial cells were incubated with PEDF the VEGF receptors showed intracellular localization. These data suggest that the interaction between PEDF and VEGFR-1 or VEGFR-2 may be a possible mechanism for inhibiting angiogenesis. PEDF may be binding to the VEGF receptors to promote their internalization and/or degradation to limit VEGF responses in treated cells.

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Acknowledgments

EKJ and JEK dedicate this study to their friend and colleague, Mary Kay Francis, who lost her battle with leukemia while the study was in progress. Mary Kay’s pioneering work on EPC-1/PEDF led to this research, and the authors treasured her enthusiasm and inspirational leadership. She is truly missed.

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Author notes

  1. Erin K. Johnston

    Present address: Janssen Research and Development, LLC, Pharma R&D Quality and Compliance, 1400 McKean Road, Building 32-12334, Spring House, PA, 19477, USA

Authors and Affiliations

  1. Department of Biology, G24A Mendel Science Center, Villanova University, 800 East Lancaster Avenue, Villanova, PA, 19085, USA

    Mary K. Francis & Janice E. Knepper

Authors
  1. Erin K. Johnston
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  2. Mary K. Francis
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  3. Janice E. Knepper
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Correspondence to Janice E. Knepper.

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Editor: T. Okamoto

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Johnston, E.K., Francis, M.K. & Knepper, J.E. Recombinant pigment epithelium-derived factor PEDF binds vascular endothelial growth factor receptors 1 and 2. In Vitro Cell.Dev.Biol.-Animal 51, 730–738 (2015). https://doi.org/10.1007/s11626-015-9884-0

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  • DOI: https://doi.org/10.1007/s11626-015-9884-0

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