regulatory elements within the prodomain of falcipain-2, a cysteine protease of the malaria parasite plasmodium falciparum监管元素falcipain-2 prodomain内,半胱氨酸蛋白酶的恶性疟原虫.pdfVIP

Regulatory Elements within the Prodomain of Falcipain- 2, a Cysteine Protease of the Malaria Parasite Plasmodium falciparum 1 2,3,4 2,4 1 Kailash C. Pandey , David T. Barkan , Andrej Sali , Philip J. Rosenthal * 1 Department of Medicine, University of California San Francisco, San Francisco, California, United States of America, 2 Departments of Biopharmaceutical Sciences and Pharmaceutical Chemistry, University of California San Francisco, San Francisco, California, United States of America, 3 Graduate Group in Bioinformatics, University of California San Francisco, San Francisco, California, United States of America, 4 California Institute for Quantitative Biosciences, University of California San Francisco, San Francisco, California, United States of America Abstract Falcipain-2, a papain family cysteine protease of the malaria parasite Plasmodium falciparum, plays a key role in parasite hydrolysis of hemoglobin and is a potential chemotherapeutic target. As with many proteases, falcipain-2 is synthesized as a zymogen, and the prodomain inhibits activity of the mature enzyme. To investigate the mechanism of regulation of falcipain-2 by its prodomain, we expressed constructs encoding different portions of the prodomain and tested their ability to inhibit recombinant mature falcipain-2. We identified a C-terminal segment (Leu155–Asp243) of the prodomain, including two motifs (ERFNIN and GNFD) that are conserved in cathepsin L sub-family papain family proteases, as the mediator of prodomain inhibitory activity. Circular dichroism analysis showed that the prodomain including the C-terminal segment, but not constructs lacking this segment, was rich in secondary structure, suggesting that the segment plays a crucial role in protein folding. Th