Expression of Novel Biomolecule for Targeting and Eradicating Intracellular Pathogens
Disciplines
Biochemistry, Biophysics, and Structural Biology | Life Sciences
Abstract (300 words maximum)
REDACT ABSTRACT AFTER ACCEPTANCE FOR IP PROTECTION. This project aims to establish a foundation for a new therapy using cell-penetrating peptides (CPPs) to deliver novel antibacterials against intracellular pathogens. This is easier said than done due to the endosomal escape problem, where molecules, in this case CPP-cargos enter the cell but remain trapped in endosomes and are either degraded or cycled back to the cell surface. We seek to address and nullify this problem by expressing our biomolecule. Rather than having the CPP directly attached to the cargo, we will use a novel non-covalent coupling strategy, a calmodulin-binding sequence (CBS) interaction with calmodulin, a calcium-dependent binder of CBS sequences. When the CPP is fused to calmodulin and the cargo, the complex of these two molecules would allow penetration into the cell and the cargo could separate from the CPP when transitioning to a low Ca2+ environment, i.e. when it enters the cell. We will be screening a multitude of cargos, with the intent that the cargo will target the pathogen of interest e.g. Staphylococcus aureus, Chlamydia trachomatis, Mycobacterium tuberculosis. We must first express the protein (with the regulatory sequence) in sufficient quantity for structural biology, enzymology, confocal microscopy, and biophysical characterization; progress toward that goal will be presented. If successful, this project may provide the groundwork for developing a new method of treating difficult intracellular bacterial pathogens.
Academic department under which the project should be listed
CSM - Chemistry and Biochemistry
Primary Investigator (PI) Name
Thomas Leeper
Expression of Novel Biomolecule for Targeting and Eradicating Intracellular Pathogens
REDACT ABSTRACT AFTER ACCEPTANCE FOR IP PROTECTION. This project aims to establish a foundation for a new therapy using cell-penetrating peptides (CPPs) to deliver novel antibacterials against intracellular pathogens. This is easier said than done due to the endosomal escape problem, where molecules, in this case CPP-cargos enter the cell but remain trapped in endosomes and are either degraded or cycled back to the cell surface. We seek to address and nullify this problem by expressing our biomolecule. Rather than having the CPP directly attached to the cargo, we will use a novel non-covalent coupling strategy, a calmodulin-binding sequence (CBS) interaction with calmodulin, a calcium-dependent binder of CBS sequences. When the CPP is fused to calmodulin and the cargo, the complex of these two molecules would allow penetration into the cell and the cargo could separate from the CPP when transitioning to a low Ca2+ environment, i.e. when it enters the cell. We will be screening a multitude of cargos, with the intent that the cargo will target the pathogen of interest e.g. Staphylococcus aureus, Chlamydia trachomatis, Mycobacterium tuberculosis. We must first express the protein (with the regulatory sequence) in sufficient quantity for structural biology, enzymology, confocal microscopy, and biophysical characterization; progress toward that goal will be presented. If successful, this project may provide the groundwork for developing a new method of treating difficult intracellular bacterial pathogens.