Trichomonas vagina/is, one of the most common sexually transmitted diseases, has been shown to increase patients' susceptibility to HIV infection and cervical cancer; moreover, resistance to metronidazole is increasing, and new drug targets must be identified in order to combat resistant strains. T vagina/is expresses cysteine proteases that have been implicated in vaginal epithelial apoptosis as well as immune system evasion. In the past the various cysteine proteases have been studied as a group, and the following work examines, one specific protease, TvCP2, in detail through Western blot analysis, immunofluorescent staining, and recombinant expression. The experiments 5 presented here suggest that aT l-CP2 over-expressing transfectant line processes CP2 and sequesters it in cellular compartments. Previous data gives strong evidence of the secretion of cysteine protease CP4 and hints at the possibility of CP2 secretion as well; however, our results show no co-localization between CP2 and CP4 in T l-CP2 over expressing transfectants, suggesting separate trafficking and different roles. To better characterize CP2 function, we attempted to express active, recombinant protein. Although Pichia pastoris serves as a reliable expression vehicle, a processing event following translation ofTvCP2 appears to have cleaved the pro-domain and, along with it, the a-secretion signal, trapping active TvCP2 within the cellular pellet. A thioreoxintagged version ofTvCP2 has been expressed in E. coli, and preliminary experiments show it may auto-activate under certain conditions, but further experimentation is required to confirm the presence of active CP2 within the fraction purified from these cells.
","abstract_html":"<p>Trichomonas vagina/is, one of the most common sexually transmitted diseases, has been shown to increase patients' susceptibility to HIV infection and cervical cancer; moreover, resistance to metronidazole is increasing, and new drug targets must be identified in order to combat resistant strains. T vagina/is expresses cysteine proteases that have been implicated in vaginal epithelial apoptosis as well as immune system evasion. In the past the various cysteine proteases have been studied as a group, and the following work examines, one specific protease, TvCP2, in detail through Western blot analysis, immunofluorescent staining, and recombinant expression. The experiments 5 presented here suggest that aT l-CP2 over-expressing transfectant line processes CP2 and sequesters it in cellular compartments. Previous data gives strong evidence of the secretion of cysteine protease CP4 and hints at the possibility of CP2 secretion as well; however, our results show no co-localization between CP2 and CP4 in T l-CP2 over expressing transfectants, suggesting separate trafficking and different roles. To better characterize CP2 function, we attempted to express active, recombinant protein. Although Pichia pastoris serves as a reliable expression vehicle, a processing event following translation ofTvCP2 appears to have cleaved the pro-domain and, along with it, the a-secretion signal, trapping active TvCP2 within the cellular pellet. A thioreoxintagged version ofTvCP2 has been expressed in E. coli, and preliminary experiments show it may auto-activate under certain conditions, but further experimentation is required to confirm the presence of active CP2 within the fraction purified from these cells.</p>","abstract_has_math":false,"creators":["Wakukawa, Christopher Keith"],"institution":null,"degree_name":"Master of Science (M.S.)","degree_level":"Thesis - Pacific Access Restricted","degree_discipline":"Biological Sciences","degree_department":null,"school":null,"contributors":["Lisa A. Wrischnik"],"advisors":[],"committee_chairs":[],"committee_members":[],"year":2012,"date_issued":"2012-01-01T08:00:00Z","date_published":"2012-01-01T08:00:00Z","updated_at":"2026-07-24T05:36:52Z","subjects":["University of the Pacific Thesis;Trichomonas vaginalis;Cysteine Proteases;Sexually transmitted diseases Pathogenesis","Life Sciences"],"languages":[],"rights":[],"rights_urls":["http://rightsstatements.org/vocab/InC/1.0/"],"identifier_entries":[]},"links":{"outbound_url":"https://scholarlycommons.pacific.edu/uop_etds/813","outbound_label":"Repository record","outbound_source":"dc:identifier"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor","label":"Contributor","values":["Lisa A. Wrischnik"]},{"key":"dc:creator","label":"Author","values":["Wakukawa, Christopher Keith"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date.available","label":"Dc Date Available","values":["2012-01-01T08:00:00Z"]},{"key":"thesis:degree_discipline","label":"Discipline","values":["Biological Sciences"]},{"key":"thesis:degree_level","label":"Degree Level","values":["Thesis - Pacific Access Restricted"]},{"key":"thesis:degree_name","label":"Degree Name","values":["Master of Science (M.S.)"]}]},{"id":"subjects_keywords","label":"Subjects and Keywords","entries":[{"key":"dc:subject","label":"Dc Subject","values":["University of the Pacific Thesis;Trichomonas vaginalis;Cysteine Proteases;Sexually transmitted diseases Pathogenesis","Life Sciences"]}]},{"id":"language_rights","label":"Language and Rights","entries":[{"key":"dc:rights","label":"Dc Rights","values":["http://rightsstatements.org/vocab/InC/1.0/"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier","label":"Identifier","values":["https://scholarlycommons.pacific.edu/uop_etds/813"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description.abstract","label":"Abstract","values":["Trichomonas vagina/is, one of the most common sexually transmitted diseases, has been shown to increase patients' susceptibility to HIV infection and cervical cancer; moreover, resistance to metronidazole is increasing, and new drug targets must be identified in order to combat resistant strains. T vagina/is expresses cysteine proteases that have been implicated in vaginal epithelial apoptosis as well as immune system evasion. In the past the various cysteine proteases have been studied as a group, and the following work examines, one specific protease, TvCP2, in detail through Western blot analysis, immunofluorescent staining, and recombinant expression. The experiments 5 presented here suggest that aT l-CP2 over-expressing transfectant line processes CP2 and sequesters it in cellular compartments. Previous data gives strong evidence of the secretion of cysteine protease CP4 and hints at the possibility of CP2 secretion as well; however, our results show no co-localization between CP2 and CP4 in T l-CP2 over expressing transfectants, suggesting separate trafficking and different roles. To better characterize CP2 function, we attempted to express active, recombinant protein. Although Pichia pastoris serves as a reliable expression vehicle, a processing event following translation ofTvCP2 appears to have cleaved the pro-domain and, along with it, the a-secretion signal, trapping active TvCP2 within the cellular pellet. A thioreoxintagged version ofTvCP2 has been expressed in E. coli, and preliminary experiments show it may auto-activate under certain conditions, but further experimentation is required to confirm the presence of active CP2 within the fraction purified from these cells.
"]},{"key":"dc:source","label":"Dc Source","values":["65"]},{"key":"dc:title","label":"Title","values":["The recombinant expression and localization of TvCP2 of trichomonas vaginalis"]}]}],"canonical_facts":{"dc:contributor":["Lisa A. Wrischnik"],"dc:creator":["Wakukawa, Christopher Keith"],"dc:date.available":["2012-01-01T08:00:00Z"],"dc:description.abstract":["Trichomonas vagina/is, one of the most common sexually transmitted diseases, has been shown to increase patients' susceptibility to HIV infection and cervical cancer; moreover, resistance to metronidazole is increasing, and new drug targets must be identified in order to combat resistant strains. T vagina/is expresses cysteine proteases that have been implicated in vaginal epithelial apoptosis as well as immune system evasion. In the past the various cysteine proteases have been studied as a group, and the following work examines, one specific protease, TvCP2, in detail through Western blot analysis, immunofluorescent staining, and recombinant expression. The experiments 5 presented here suggest that aT l-CP2 over-expressing transfectant line processes CP2 and sequesters it in cellular compartments. Previous data gives strong evidence of the secretion of cysteine protease CP4 and hints at the possibility of CP2 secretion as well; however, our results show no co-localization between CP2 and CP4 in T l-CP2 over expressing transfectants, suggesting separate trafficking and different roles. To better characterize CP2 function, we attempted to express active, recombinant protein. Although Pichia pastoris serves as a reliable expression vehicle, a processing event following translation ofTvCP2 appears to have cleaved the pro-domain and, along with it, the a-secretion signal, trapping active TvCP2 within the cellular pellet. A thioreoxintagged version ofTvCP2 has been expressed in E. coli, and preliminary experiments show it may auto-activate under certain conditions, but further experimentation is required to confirm the presence of active CP2 within the fraction purified from these cells.
"],"dc:identifier":["https://scholarlycommons.pacific.edu/uop_etds/813"],"dc:rights":["http://rightsstatements.org/vocab/InC/1.0/"],"dc:source":["65"],"dc:subject":["University of the Pacific Thesis;Trichomonas vaginalis;Cysteine Proteases;Sexually transmitted diseases Pathogenesis","Life Sciences"],"dc:title":["The recombinant expression and localization of TvCP2 of trichomonas vaginalis"],"thesis:degree_discipline":["Biological Sciences"],"thesis:degree_level":["Thesis - Pacific Access Restricted"],"thesis:degree_name":["Master of Science (M.S.)"]},"updated_at":"2026-07-24T05:36:52Z"}