Discipline: Biological Sciences
Subcategory: Cell and Molecular Biology
Raveen Hough - University of the District of Columbia
Co-Author(s): Carolyn Cousin and Matty Knight, University of the District of Columbia Michael Smith, Howard University
Schistosomiasis is a major public health concern in the tropics and sub-tropics. The disease is transmitted by a parasitic trematode that requires a fresh water snail for the development of the larval forms of the parasite. The asexual development of the intra-molluscan stages of the parasite produces the infectious cercariae that penetrate human skin in order to gain access in the blood stream where they develop into the other stages (schistosomula, and adult worms) of the parasites life cycle. There is only one effective drug (praziquantel) to treat schistosomiasis, and no preventative vaccine. Use of a mass chemotherapy approach to control the spread of the disease has been challenging. With global warming, schistosomiasis has now emerged in Southern Europe. There are now calls for new alternative control strategies that are based on eliminating the infection in the snail host. Previously, it was shown that in the snail Biomphalaria glabrata and its compatible schistosome species, Schistosoma mansoni, that an early stress response in the snail host is a necessary step for snail susceptibility. Stress proteins Hsp70 and Hsp90 were induced within the first 30 minutes following infection of the NMRI susceptible but not the resistant BS90 snail. In order to identify signaling pathways that enable the snail to accommodate parasite development that can be interfered with to block transmission, we have used a co-immunoprecipitation approach to identify proteins that are closely associated with one of the major stress proteins, Hsp70. Using mono-specific antibodies raised against the snail recombinant Hsp70, soluble and insoluble (membrane bound) protein complexes were captured on protein A- Sepharose beads that were further analyzed by SDS-PAGE. Results showed that the snail Hsp70 cognate protein associates with multiple client proteins ranging from molecular weight > 200 to 14kDa. To reduce the number of Hsp70 bound proteins, the recombinant protein has been purified and will be labeled with biotin and used as bait to only capture client proteins that can be affinity purified on a Streptavidin- agarose beads.
Funder Acknowledgement(s): Freddie Dixon, Anita Wood, Carolyn Cousin, and Matty Knight, University of the District of Columbia.
Faculty Advisor: Matty Knight, Matty_Knight@email.gwu.edu
Role: I was responsible for the co-immunoprecipitation process, maintenance of the snails used, and identifying what client proteins were attached using SDS protein gels.