Discipline: Chemistry and Chemical Sciences
Subcategory: Biochemistry (not Cell and Molecular Biology and Genetics)
Erica Brackett - Dalton State College
Co-Author(s): Matt C. McInturff, Kristen V. Coutinho, and Carol A. Chrestensen, Kennesaw State University, Kennesaw, GA
If TAT-CaM can deliver proteins to a single cell, then it will be able to deliver proteins to a population of cells. Being able to deliver proteins to a population of cells can be used in many medical applications, such as delivering lymphocyte proteins to a population of cancer cells. Intracellular delivery of proteins to cells has always had the obstacle of getting through the cell membrane. Cell penetrating peptides (CPPs) have been shown to get through the membrane of various cell types and enter cells. Our novel CPP with the ability to release cargo, has yet to be shown to impact a population of cells. Our recent results have shown that TAT-CaM has been shown to deliver non-covalently bound cargos into cells utilizing the calmodulin/calmodulin binding protein interactions. Our experiments were designed to determine the ability of TAT-CaM to deliver cargo efficiently into a population of cells; all our previous experiments have relied on single cell measurements. The cargos used in this experiment were CBP-Myoglobin and CBP-p38. These experiments tested different types of cells, such as C2C12 myoblasts, S2 cells, and BHK cells, for the ability to modify a population of cells. Our results seem to show that TAT-CaM is leaving the cargo in the membrane of the cells. We would like to know why this happened and have several hypotheses, such as that the amount of calcium in the lysis buffer may be causing the TAT-CaM to bind back to the cargo and the membrane. Further research will be focused on using different amounts of calcium in the lysis buffer to see how that effects the ability of TAT-CaM to get cargo into the cells of a population.
Funder Acknowledgement(s): NSF REU KSU CBSURE #431408; NIH R15GM110634.
Faculty Advisor: Carol Chrestensen, email@example.com
Role: This was my first research experience and I found it to be quite educating. Within the first two weeks I was shown how to operate most of the lab equipment I would need and how to preform various tasks using sterile technique. By the third week I was running supervised experiments and soon I was even allowed to plan out my own experiments. I preformed all of the cell penetration assays myself and most of the western blots. With Dr. Chrestensen's help I analysed the results of my western blots and created the plans for following experiments based upon what questions I wanted to test.