Title

Characterization of an Adenosylhomocysteine Hydrolase Enzyme in the protozoal parasite Trichomonas vaginalis

Poster Number

44

Lead Author Major

Biological Sciences

Format

Poster Presentation

Faculty Mentor Name

Kirkwood Land

Faculty Mentor Department

Biological Sciences

Abstract/Artist Statement

Trichomonas vaginalis is the causative agent of trichomoniasis, a common sexually-transmitted disease in humans. Approximately 5% of cases of trichomoniasis are resistant to treatment with the commonly prescribed metronidazole. The search for alternative new therapies for both nitroimidazole susceptible and resistant cases is imperative. Here, we have shown that 2'-deoxy- 2'-fluoroadenosine, 9-(β,D- arabinofuranosyl)adenine, 9-(2-deoxy-2-fluoro- β,D-arabinofuranosyl)adenine , and 9-(2-chloro- 2-deoxy-β,D-arabinofuranosyl)adenine inhibit T. vaginalis 100% at 100 μM level. These compounds have an IC50 of 2.94 μM, 3.6 μM, 0.09 μM, and 5.93 μM, respectively (Metronidazole’s IC50 value for the same strain is 0.72 μM). To further characterize this potential drug target, we have analyzed the ADHY activity in whole cell extracts of T. vaginalis. We have also cloned the gene into pQE80L and have successfully expressed the enzyme in E.coli. To characterize the subcellular location of this enzyme, we have constructed an expression vector with the gene for an HA- epitope tag on the C-terminus of the ADHY protein. The successful expression, characterization, and localization of the ADHY protein of T. vaginalis will set the stage for development of inhibitors against this enzyme as chemotherapy for drug resistant infections.

Location

Grave Covell

Start Date

21-4-2012 10:00 AM

End Date

21-4-2012 12:00 PM

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Apr 21st, 10:00 AM Apr 21st, 12:00 PM

Characterization of an Adenosylhomocysteine Hydrolase Enzyme in the protozoal parasite Trichomonas vaginalis

Grave Covell

Trichomonas vaginalis is the causative agent of trichomoniasis, a common sexually-transmitted disease in humans. Approximately 5% of cases of trichomoniasis are resistant to treatment with the commonly prescribed metronidazole. The search for alternative new therapies for both nitroimidazole susceptible and resistant cases is imperative. Here, we have shown that 2'-deoxy- 2'-fluoroadenosine, 9-(β,D- arabinofuranosyl)adenine, 9-(2-deoxy-2-fluoro- β,D-arabinofuranosyl)adenine , and 9-(2-chloro- 2-deoxy-β,D-arabinofuranosyl)adenine inhibit T. vaginalis 100% at 100 μM level. These compounds have an IC50 of 2.94 μM, 3.6 μM, 0.09 μM, and 5.93 μM, respectively (Metronidazole’s IC50 value for the same strain is 0.72 μM). To further characterize this potential drug target, we have analyzed the ADHY activity in whole cell extracts of T. vaginalis. We have also cloned the gene into pQE80L and have successfully expressed the enzyme in E.coli. To characterize the subcellular location of this enzyme, we have constructed an expression vector with the gene for an HA- epitope tag on the C-terminus of the ADHY protein. The successful expression, characterization, and localization of the ADHY protein of T. vaginalis will set the stage for development of inhibitors against this enzyme as chemotherapy for drug resistant infections.