Biomphalaria glabrata snails are known to display a wide range of susceptibility phenotypes to Schistosoma mansoni infection depending on the genetics of both the snail and the invading parasite. Evidence exists for a role of hydrolytic enzymes in the defense of molluscs against invading parasites. To elucidate the role of these enzymes in the outcome of infection in the snail, proteolysis was examined in parasite-resistant and -susceptible snails. Zymographs of extracts from the whole snail or hepatopancreas indicated higher proteolytic activity in resistant, compared with susceptible, snails. Lytic activity coincided with a high-molecular-weight smear (220 to 66 kDa) that was abrogated by the cysteine protease inhibitor trans-epoxysuccinyl-l-leucylamido-(4-guanidino)butane. Quantitative flourimetric assays showed 3.5-fold higher activity in resistant than in susceptible snails. From a hepatopancreas cDNA library, several cysteine protease encoding expressed sequence tags including the full-length cDNA for cathepsin B were identified. Sequence analysis revealed that this cathepsin B belonged to the C1A family of peptidases characterized by the presence of the catalytic cysteine–histidine dyad, the “occluding loop,” signal sequence, and cleavage sites for the prepro and propeptides. Quantitative real-time reverse transcriptase-polymerase chain reaction showed higher up-regulation of cathepsin B transcript in resistant than in the susceptible snail after parasite exposure.
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June 2008
INVERTEBRATE-PARASITE RELATIONSHIPS|
June 01 2008
Differences in Cysteine Protease Activity in Schistosoma mansoni-Resistant and -Susceptible Biomphalaria glabrata and Characterization of the Hepatopancreas Cathepsin B Full-length cDNA
Jocelyn Myers;
Jocelyn Myers
Biomedical Research Institute, 12111 Parklawn Drive, Rockville, Maryland 20852. mknight@afbr-bri.com
* Department of Biology, Howard University, 2400 Sixth Street NW, Washington, D.C. 20059
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Wannaporn Ittiprasert;
Wannaporn Ittiprasert
Biomedical Research Institute, 12111 Parklawn Drive, Rockville, Maryland 20852. mknight@afbr-bri.com
† Institute of Science and Technology for Research and Development, Mahidol University, Salaya Campus, Nakhon Pathom, Thailand 73170
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Nithya Raghavan;
Nithya Raghavan
Biomedical Research Institute, 12111 Parklawn Drive, Rockville, Maryland 20852. mknight@afbr-bri.com
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André Miller;
André Miller
Biomedical Research Institute, 12111 Parklawn Drive, Rockville, Maryland 20852. mknight@afbr-bri.com
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Matty Knight
Matty Knight
Biomedical Research Institute, 12111 Parklawn Drive, Rockville, Maryland 20852. mknight@afbr-bri.com
‡ To whom correspondence should be addressed
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J Parasitol (2008) 94 (3): 659–668.
Citation
Jocelyn Myers, Wannaporn Ittiprasert, Nithya Raghavan, André Miller, Matty Knight; Differences in Cysteine Protease Activity in Schistosoma mansoni-Resistant and -Susceptible Biomphalaria glabrata and Characterization of the Hepatopancreas Cathepsin B Full-length cDNA. J Parasitol 1 June 2008; 94 (3): 659–668. doi: https://doi.org/10.1645/GE-1410.1
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