Project Title

Does CPP delivered catalase have measurable physiological effects?

Presenters

Academic department under which the project should be listed

CSM - Molecular and Cellular Biology

Faculty Sponsor Name

Susan Smith

Additional Faculty

Jonathan McMurry, Biology, jmcmurr1@kennesaw.edu Julia LeCher, Biology, jwand@kennesaw.edu

Abstract (300 words maximum)

Catalase is an enzyme found in nearly all living organisms and is used as a defense against oxidative stress, specifically by degrading the hydrogen peroxide (H2O2) into water and oxygen. We have demonstrated successful penetration of catalase into the BHK mammalian cells using cell penetrating peptide (CPP) technology: both fluorescence from catalase conjugated with amino dye 555, and catalase activity in cell lysates, is significantly higher in CPP+catalase treated cells than in cells treated with catalase alone. Here we examine two questions related to these results. First, does the catalase traffic to peroxisomes, its correct location in mammalian cells? Secondly, does catalase delivered as cargo on a cell penetrating peptide protect mammalian cells against damage from H2O2? Confocal microscopy was used to quantify the colocalization of TaT-CaM delivered catalase to the peroxisome, the known subcellular location of catalase, in baby hamster kidney (BHK) cells. Cell viability was assessed by treating BHK cells with different concentrations of hydrogen peroxide for different times. Protein S-glutathionylation provided a method for assessing physiological effects of CPP delivered catalase. BHK cells exposed to H2O2 showed increased levels of S-glutathionylation of proteins. Preliminary densitometry analysis shows a statistically significant difference between the S-glutathionylation in CPP catalase treated cells and the non treated cells. Here, we present results from colocalization and physiological protection of CPP delivered catalase assayed to date.

Project Type

Oral Presentation (15-min time slots)

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Does CPP delivered catalase have measurable physiological effects?

Catalase is an enzyme found in nearly all living organisms and is used as a defense against oxidative stress, specifically by degrading the hydrogen peroxide (H2O2) into water and oxygen. We have demonstrated successful penetration of catalase into the BHK mammalian cells using cell penetrating peptide (CPP) technology: both fluorescence from catalase conjugated with amino dye 555, and catalase activity in cell lysates, is significantly higher in CPP+catalase treated cells than in cells treated with catalase alone. Here we examine two questions related to these results. First, does the catalase traffic to peroxisomes, its correct location in mammalian cells? Secondly, does catalase delivered as cargo on a cell penetrating peptide protect mammalian cells against damage from H2O2? Confocal microscopy was used to quantify the colocalization of TaT-CaM delivered catalase to the peroxisome, the known subcellular location of catalase, in baby hamster kidney (BHK) cells. Cell viability was assessed by treating BHK cells with different concentrations of hydrogen peroxide for different times. Protein S-glutathionylation provided a method for assessing physiological effects of CPP delivered catalase. BHK cells exposed to H2O2 showed increased levels of S-glutathionylation of proteins. Preliminary densitometry analysis shows a statistically significant difference between the S-glutathionylation in CPP catalase treated cells and the non treated cells. Here, we present results from colocalization and physiological protection of CPP delivered catalase assayed to date.