Possible anti-angiogenic activity of the recombinant mojastin mutant disintegrin, r-Moj-MP

Integrins, transmembrane proteins that act as cell-matrix adhesion receptors, plays an important role in the process of angiogenesis, the formation of new blood vessels. Disintegrins, found in viper venom, are small peptides containing an arginine (R), glycine (G) and Aspartate (D) binding motif that have a high binding affinity to integrins. The recombinant version of the disintegrin mojastin (r-Moj-WN) was mutated to contain the amino acids Methionine (M) and Proline (P) carboxyl to the RGD motif.  Previous research have shown that disintegrins with a RGDMP motif inhibit angiogenesis. We will be investigating the effects of r-Moj-MP peptide on tube formation in vitro. Our hypothesis is that r-Moj-MP will inhibit angiogenesis. We will be using the in vitro angiogenesis assay with cultured Human Umbilical Vein Epithelial Cells, HUVEC, to test vessel tube formation in the presence of r-Moj-MP.  Our experimental approach will involve the determination of the minimum concentration of r-Moj-MP peptide required to inhibit vessel tube formation. Thus, initially we will incubate cells with r-Moj-MP peptide at 1 µM, 2 µM, and 5µM. Previous research have shown that other disintegrins inhibit vessel tube formation at 2 µM.  So far we have expressed and purified r-Moj-MP peptide and we will begin the angiogenesis assay shortly. We predict to obtain a significantly lower amount of vessel tube formation in wells containing r-Moj-MP. This project was funded by NSF-REU DBI 1004530.