Engineering Perfringolysin O Mutants with Different Cholesterol Binding Thresholds

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dc.contributorBrowne, Sheila
dc.contributorCamp, Amy
dc.contributor.advisorKnight, Jeff
dc.contributor.advisorHeuck, Alejandro
dc.contributor.authorMote, Francisca Sefakor
dc.date.accessioned2013-05-16T00:32:25Z
dc.date.available2013-05-16T00:32:25Z
dc.date.gradyear2013en_US
dc.date.issued2013-05-15
dc.description.abstractCholesterol is a sterol molecule that is a primary constituent of mammalian cells. It is important for controlling membrane fluidity, stability and organization. It is also precursor for the biosynthesis of steroid hormones and bile acids. Abnormal levels of cholesterol have been implicated in some cardiac and brain conditions such as coronary heart disease. Perfringolysin O (PFO) is a water-soluble toxin, whose binding and insertion into cell membranes requires cholesterol. Cholesterol activity in a membrane is related to the ability of cholesterol to interact with molecules at the membrane surface. The binding of PFO is thought to be dependent on cholesterol activity and requires high cholesterol levels on the membrane’s surface for binding. Currently, cholesterol probes, such as filipin, measure overall cholesterol concentration whereas PFO measures cholesterol activity. The aim of this project is to design a probe using PFO that would detect a wide range of cholesterol activity levels. Based on previous research, we hypothesized that hydrophobic mutations in the non-conserved regions of PFO would allow the protein to interact better with the hydrophobic membrane core, and thus require less cholesterol. The additive effect of two previously identified mutants, D434S and A437V was also determined. A401L and A401F required less cholesterol compared to native PFO. The double mutant did not display an additive effect.en_US
dc.description.sponsorshipBiochemistryen_US
dc.identifier.urihttp://hdl.handle.net/10166/3233
dc.language.isoen_USen_US
dc.rights.restrictedrestricteden_US
dc.subjectCholesterolen_US
dc.subjectPerfringolysin Oen_US
dc.titleEngineering Perfringolysin O Mutants with Different Cholesterol Binding Thresholdsen_US
dc.typeThesis
mhc.degreeUndergraduateen_US
mhc.institutionMount Holyoke College

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