10.1371/journal.pntd.0001398.g003 Manuel Castro de Moura Manuel Castro de Moura Francesc Miro Francesc Miro Jung Min Han Jung Min Han Sunghoon Kim Sunghoon Kim Antonio Celada Antonio Celada Lluís Ribas de Pouplana Lluís Ribas de Pouplana EhKRS protease processing. Public Library of Science 2011 protease 2011-11-29 02:18:08 Figure https://plos.figshare.com/articles/figure/_EhKRS_protease_processing_/378288 <p>Immunoblots analysis with α-EELP antibody of recombinant EhKRS protein digestions using <i>Entamoeba</i> lysates (A) or human leukocyte elastase (B). (A) 1 µg recombinant EhKRS was incubated at 37°C for 30 minutes without <i>E. histolytica</i> crude extract (Eh CE; lane 1) or with 0.5 µg Eh CE (lane 2–4); 1 µg Eh CE (lane 5–7); 5 µg Eh CE (lane 8–10) or 5 µg Eh CE plus protease inhibitors (lane 11–13) for 5, 15 or 30 minutes. (B) Digestion of 1 µg recombinant EhKRS protein with elastase for 30 minutes at 37°C. (C) EhMRS protease processing. Digestion of 1 µg recombinant EhMRS at 5, 15 and 30 minutes with 1 µg of Eh CE (lane 2–4) or 5 µg Eh CE (lane 5–7). Recombinant EhMRS control without Eh CE (lane 1). Digestion products were detected by immunoblot using α-EELP antibody. Arrow shows EELP product resulting from recombinant EhKRS or EhMRS. Asterisk denotes recombinant EELP with 6 His tag plus 36 amino acids at the N-terminal. Boxed sequence corresponds to the N-terminal sequence of EELP after digestion from EhKRS by elastase, as determined by Edman degradation.</p>