Proteinases in Excretory-Secretory Products of Toxocara canis Second-Stage Larvae: Zymography and Modeling Insights

Figure 2

Proteolytic activity of TES components of T. canis analyzed by acrylamide-gelatin gel electrophoresis. Effect of proteinase inhibitors. TES components were preincubated for 30 min at RT with one of the following compounds: control without inhibitor (lane 1); 2 μM trans-epoxysuccinyl-L-leucylamido(4-guanidino)-butane (E64) to identify cysteine proteinases (lane 2); 0.1 μM pepstatin A for aspartic proteinases (lane 3); 1 mM ethylenediaminetetraacetic acid (EDTA) for metalloproteinases (lane 4); and 0.1 μM leupeptin for serine proteinases (lane 5). Samples (10 μg/well) were electrophoretically separated in slab gels of 10% (w/v) acrylamide copolymerized with 0.1% (w/v) gelatin. Proteinase activity was developed using phosphate buffer (pH 7.6) and then gels were stained with Coomassie blue. Molecular weight of bands with activity is indicated at the right.