Discipline: Biological Sciences
Subcategory: Cell and Molecular Biology
India El - Norfolk State University
Co-Author(s): Brittney K. Burton, Norfolk State University, Norfolk, VA
Calcium ions (Ca2+) and Protein Kinase C enzymes are necessary for the release of insulin. The calcium ions stimulate or fuel the release of insulin into the bloodstream. In previous works it has been discovered that Beta (β) subunit blocks insulin secretion and the mechanism is not known. It is hypothesized that Beta (β) subunit interferes with the interaction between the calcium channel Alpha 1 (α1) subunit and the PKC enzymes. The purpose of this research is to understand this interaction between the subunit proteins and determine the basis of the inhibition of insulin secretion by β subunit. Cav Alpha 1 (α1) and Beta (β) subunit proteins were synthesized in the lab. Concentrations of β3 proteins were obtained; Sample 1 (β3 insert +PFN21K) 1.206mg/mL, Sample 2 (β3 insert +PFN21K) 1.159mg/mL, Sample 3 (β3 insert +PFN21K) 1.3735mg/mL, Sample 4 (PFN21K-Control) 1.3335mg/mL. Protein concentrations have been obtained which will be analyzed through the SDS-PAGE and further analyzed through Western Blotting. SDS-PAGE and Western Blotting will be used to reassure that we have the appropriate protein size. Obtaining the proteins from the DNA will allow us to further the research by combining the α1 and β in-vitro and studying their interaction. This combination of the proteins will expectantly yield the location of the blockage Beta creates on the Alpha 1 subunit.
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Funder Acknowledgement(s): I thank Ganesan Kamatchi and Brittney K. Burton for the guidance with this experiment. I also thank Zenora Spellman for the opportunity to participate in the research experience. Funding was provided by NSU/HBCU-UP.
Faculty Advisor: Ganesan Kamatchi, email@example.com
Role: I did each part of the experiment which includes PCR Reaction, electrophoresis gel analysis, gel extraction, bacterial transformation(Eschericia coli or E.Coli), transfections (HEK-293 human cells), lysis of mammalian cells, and protein estimations.