The recombinant expression and localization of TvCP2 of trichomonas vaginalis
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 strain...
Main Author: | |
---|---|
Format: | Others |
Published: |
Scholarly Commons
2012
|
Subjects: | |
Online Access: | https://scholarlycommons.pacific.edu/uop_etds/813 https://scholarlycommons.pacific.edu/cgi/viewcontent.cgi?article=1812&context=uop_etds |
id |
ndltd-pacific.edu-oai-scholarlycommons.pacific.edu-uop_etds-1812 |
---|---|
record_format |
oai_dc |
spelling |
ndltd-pacific.edu-oai-scholarlycommons.pacific.edu-uop_etds-18122021-10-05T05:13:17Z The recombinant expression and localization of TvCP2 of trichomonas vaginalis Wakukawa, Christopher Keith 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. 2012-01-01T08:00:00Z text application/pdf https://scholarlycommons.pacific.edu/uop_etds/813 https://scholarlycommons.pacific.edu/cgi/viewcontent.cgi?article=1812&context=uop_etds University of the Pacific Theses and Dissertations Scholarly Commons University of the Pacific Thesis;Trichomonas vaginalis;Cysteine Proteases;Sexually transmitted diseases Pathogenesis Life Sciences |
collection |
NDLTD |
format |
Others
|
sources |
NDLTD |
topic |
University of the Pacific Thesis;Trichomonas vaginalis;Cysteine Proteases;Sexually transmitted diseases Pathogenesis Life Sciences |
spellingShingle |
University of the Pacific Thesis;Trichomonas vaginalis;Cysteine Proteases;Sexually transmitted diseases Pathogenesis Life Sciences Wakukawa, Christopher Keith The recombinant expression and localization of TvCP2 of trichomonas vaginalis |
description |
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. |
author |
Wakukawa, Christopher Keith |
author_facet |
Wakukawa, Christopher Keith |
author_sort |
Wakukawa, Christopher Keith |
title |
The recombinant expression and localization of TvCP2 of trichomonas vaginalis |
title_short |
The recombinant expression and localization of TvCP2 of trichomonas vaginalis |
title_full |
The recombinant expression and localization of TvCP2 of trichomonas vaginalis |
title_fullStr |
The recombinant expression and localization of TvCP2 of trichomonas vaginalis |
title_full_unstemmed |
The recombinant expression and localization of TvCP2 of trichomonas vaginalis |
title_sort |
recombinant expression and localization of tvcp2 of trichomonas vaginalis |
publisher |
Scholarly Commons |
publishDate |
2012 |
url |
https://scholarlycommons.pacific.edu/uop_etds/813 https://scholarlycommons.pacific.edu/cgi/viewcontent.cgi?article=1812&context=uop_etds |
work_keys_str_mv |
AT wakukawachristopherkeith therecombinantexpressionandlocalizationoftvcp2oftrichomonasvaginalis AT wakukawachristopherkeith recombinantexpressionandlocalizationoftvcp2oftrichomonasvaginalis |
_version_ |
1719487481503547392 |