Journal: Toxins

Article Title: A Novel P-III Metalloproteinase from Bothrops barnetti Venom Degrades Extracellular Matrix Proteins, Inhibits Platelet Aggregation, and Disrupts Endothelial Cell Adhesion via α5β1 Integrin Receptors to Arginine–Glycine–Aspartic Acid (RGD)-Containing Molecules

doi: 10.3390/toxins16110486

Figure Lengend Snippet: ( A ) Western blot analysis of Bar-III on recombinant vWF-A1 domain (rvWF-A1). vWF-A1 (5 µg) was treated with Bar-III (4 µg) for 60 min at 37 °C, as described in methodology section. The proteolysis of vWF-A1 (indicated by an arrow) was analyzed using SDS-PAGE (7–15% gradient gel) and blotted with mice anti-vWF IgG. ( B ) Bar-III does not cleave the vWF receptor partner GPIbα on platelets. Washed platelets were incubated with Bar-III (4 µg) at 37 °C at the indicated intervals, and the reactions were terminated by addition of SDS loading sample buffer. The platelet lysate was probed in WB with anti-CD42/GPIb. Note the intact ~130-kDa GPIbα expression on platelets. ( C ) Platelet pellets or supernatants of WPs treated with Bar-III (4 µg) for 60 min at 37 °C. The positions of GPVI (~62-kDa) and the soluble GPVI fragment (~55-kDa) are indicated. C, platelet control. These results are representative of at least three similar experiments for each item.

Article Snippet: The GPVI polyclonal antibody (AF3627) of human platelets and CD42b/GPIbα (antihuman) polyclonal antibody (AF4067) were acquired from R&D Systems (Minneapolis, MN, USA).

Techniques: Western Blot, Recombinant, SDS Page, Incubation, Expressing, Control