Infection by parasitic helminthes is a large health issue that causes many health problems for people globally, particularly in the third world. Although drugs exist for the infection of these parasites, many cause negative side effects for the host and many of the worms are developing resistance to the drugs. It is therefore important to discover new drug targets that are less harmful to the host and which the parasites are not growing a resistance toward. While in the host these parasitic worms use an anaerobic metabolic pathway due to the lack of oxygen in their environment. R. rubrum also uses an anaerobic pathway in low oxygen conditions that is similar to that of the helminthes. Rhodoquinone (RQ) is an electron carrier that is needed for the anaerobic respiration of R. rubrum. RquA is a  protein that was recently discovered to be required for the synthesis of RQ in R. rubrum. RquA is a good potential drug target as RQ is not found in mammalian species which solely use an aerobic pathway for respiration. An overexpression method for RquA will be optimized using a H2 mutant strain of R. rubrum order tooverexpress RquA. RquA will be purified using nickel affinity chromatography as well as size exclusion chromatography. A SAM kinetic assay will be used in order to confirm the belief that RquA is a Sam dependent methyl-transferase. X-ray crystallography will also be performed on RquA in order to find the tertiary structure as well as be able to determine the active site and binding residues.  These studies on the activity of RquA should help in future attempts to make an inhibitor for RquA which could potentially be used in a novel drug.

Purification and characterization of RquA

247th National Meeting (2014)

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