CPP-Mediated Delivery of Peptidoglycan Hydrolases as Novel Enzybiotics

Disciplines

Biochemistry | Cell Biology | Molecular Biology

Abstract (300 words maximum)

CPP-Mediated Delivery of Peptidoglycan Hydrolases as Novel Enzybiotics

Jacob Clawson, Tina Willoughby, Daniel Morris and Jonathan McMurry Department of Biochemistry, Kennesaw State University

In the medical field there has always been a topic of discussion on how to deal with nosocomial diseases. Recently there has been an upturn of pathogens that are nosocomial while also being antibiotic resistant as they hide inside the cell. Peptidoglycan hydrolases (PHs), which degrade the cell wall structures of bacteria, are a promising alternative to traditional chemotherapeutics. However, against intracellular pathogens such as Staphylococcus aureus, they, too, face a significant problem in crossing the cell membrane. Cell-penetrating peptides (CPPs) offer the possibility of overcoming delivery barriers for biomolecules, but protein ‘cargos’ often get trapped in the endosomal pathway, failing to escape which ultimately averts the cargo from getting to its desired destination. The McMurry Group developed TAT-CaM to solve the problem by utilizing non-covalent, Ca2+ -dependent coupling between CCP to a cargo via calmodulin-calmodulin binding site (CBS) interactions. By employing TAT-CaM to deliver PHs with engineered CBSs, we hypothesize that we will be able to efficiently deliver PHs to mammalian cell interiors, resolving intracellular infections. In this study, several CBS-PHs were designed, expressed, purified, characterized and delivered to mammalian BHK cells. First steps in assaying antibacterial activity in a tissue culture infection model will also be described. Success will enable further initial development of a novel, a class of antibacterial therapeutics.

Academic department under which the project should be listed

CSM - Molecular and Cellular Biology

Primary Investigator (PI) Name

Jonathan McMurry

Additional Faculty

Daniel Morris, Molecular and Cellular Biology, dmorr125@kennesaw.edu

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CPP-Mediated Delivery of Peptidoglycan Hydrolases as Novel Enzybiotics

CPP-Mediated Delivery of Peptidoglycan Hydrolases as Novel Enzybiotics

Jacob Clawson, Tina Willoughby, Daniel Morris and Jonathan McMurry Department of Biochemistry, Kennesaw State University

In the medical field there has always been a topic of discussion on how to deal with nosocomial diseases. Recently there has been an upturn of pathogens that are nosocomial while also being antibiotic resistant as they hide inside the cell. Peptidoglycan hydrolases (PHs), which degrade the cell wall structures of bacteria, are a promising alternative to traditional chemotherapeutics. However, against intracellular pathogens such as Staphylococcus aureus, they, too, face a significant problem in crossing the cell membrane. Cell-penetrating peptides (CPPs) offer the possibility of overcoming delivery barriers for biomolecules, but protein ‘cargos’ often get trapped in the endosomal pathway, failing to escape which ultimately averts the cargo from getting to its desired destination. The McMurry Group developed TAT-CaM to solve the problem by utilizing non-covalent, Ca2+ -dependent coupling between CCP to a cargo via calmodulin-calmodulin binding site (CBS) interactions. By employing TAT-CaM to deliver PHs with engineered CBSs, we hypothesize that we will be able to efficiently deliver PHs to mammalian cell interiors, resolving intracellular infections. In this study, several CBS-PHs were designed, expressed, purified, characterized and delivered to mammalian BHK cells. First steps in assaying antibacterial activity in a tissue culture infection model will also be described. Success will enable further initial development of a novel, a class of antibacterial therapeutics.