a fret-based high throughput screening assay to identify inhibitors of anthrax protective antigen binding to capillary morphogenesis gene 2 proteinfret-based高通量筛选试验来确定抑制剂炭疽的保护性抗原结合毛细管形态发生蛋白基因2.pdfVIP

A FRET-Based High Throughput Screening Assay to Identify Inhibitors of Anthrax Protective Antigen Binding to Capillary Morphogenesis Gene 2 Protein 1 1 1 1 2 Michael S. Rogers , Lorna M. Cryan , Kaiane A. Habeshian , Lauren Bazinet , Thomas P. Caldwell , P. 2 2 Christine Ackroyd , Kenneth A. Christensen * 1 Department of Surgery, Vascular Biology Program, Children’s Hospital Boston, Harvard Medical School, Boston, Massachusetts, United States of America, 2 Department of Chemistry, Clemson University, Clemson, South Carolina, United States of America Abstract Anti-angiogenic therapies are effective for the treatment of cancer, a variety of ocular diseases, and have potential benefits in cardiovascular disease, arthritis, and psoriasis. We have previously shown that anthrax protective antigen (PA), a non- pathogenic component of anthrax toxin, is an inhibitor of angiogenesis, apparently as a result of interaction with the cell surface receptors capillary morphogenesis gene 2 (CMG2) protein and tumor endothelial marker 8 (TEM8). Hence, molecules that bind the anthrax toxin receptors may be effective to slow or halt pathological vascular growth. Here we describe development and testing of an effective homogeneous steady-state fluorescence resonance energy transfer (FRET) high throughput screening assay designed to identify molecules that inhibit binding of PA to CMG2. Molecules identified in the screen can serve as potential lead compounds for the development of anti-angiogenic and anti-anthrax therapies. The assay to screen for inhibitors of this protein–protein interaction is sensitive and robust, with observed Z’ values as high as 0.92. Preliminar