Natalie D’Antonio
Expression of the Human Leutenizing Hormone Receptor in E. coli
Human leutenizing hormone receptor (hLHR) is a G-protein coupled receptor endogenous to gonadal tissue involved in the regulation of steroidogenesis and gonadal cell maturation and function. Inappropriate expression of hLHR in the adrenal gland results in hypercortisolemia. It is hypothesized that the development and administration of an anti-hLHR antibody is an alternative route for the interruption of LH induced hypercortisolemia. A portion of the hLHR protein was expressed from a plasmid vector for subsequent purification. Partial purification of the protein has been completed. Additonal purification is still required before the protein can be used to generate an anti-hLHR antibody for future trials.
Jen Paludi
Analysis of an herbal supplement reveals caffeine as the primary component
Chemical characterization of herbal supplements allows for better understanding of the active molecules in those extracts and can aid in the design of better drugs for their specific targets. To characterize an over-the-counter herbal dietary supplement claiming to inhibit cortisol production, capsules were extracted into different solvents and analyzed via GC/MS. In the water extract, the most abundant ingredient detected was caffeine, which represented 98.7% of the total water- soluble compounds. In acetonitrile, the most prominent ingredient was equilenin, a steroid-like molecule. Extraction in hexane and other non-polar solvents revealed the presence of several sterol derivatives including campesterol, stigmasterol, and sitosterol. The manufacturers state that the main ingredients originate from Magnolia officinalis, the bark of magnolia. It is possible that equilenin detected in the acetonitrile extract is magnolol or honokiol, the “active ingredients” of magnolia bark extract. Further testing is needed to see how these compounds might inhibit cortisol synthesis.
Andrea Panczykowski
Ligand Independent Phosphorylation of the Human Follicle Stimulating Hormone Receptor
The purpose of this study was to investigate ligand-independent phosphorylation of the human follicle stimulating hormone receptor (hFSHR). The ligand for this receptor is the follicle stimulating hormone which is primarily responsible for oocyte development in females and spermatogenesis in males. The human follicle stimulating hormone receptor is an example of a G-protein coupled receptor (GPCR). Ligand-independent phosphorylation is hypothesized to be a mechanism for preventing activation of a GPCR, such as hFSHR. To investigate this hypothesis, a truncated hFSHR incapable of ligand binding was designed and constructed using recombinant DNA technology. Future studies will include transfection of cells with the truncated receptor to test for expression of the designed receptor on the cell surface.
Laura Pittenger
Clarifying the Mechanism of Anti-Oncotic activity of an Alpha-fetoprotein derived peptide in estrogen dependent human breast cancer
This study inquires into the mechanism of interaction on a cellular level, of a peptide derived from alpha-fetoprotein known as growth inhibitor peptide (GIP). This peptide is known to inhibit growth of estrogen dependent breast cancer in both humans and mice. We hypothesized that GIP interacts with a membrane receptor on breast cancers to carry out its activity. To test this, we cultured lines of MCF-7 human breast cancer cells and incubated these cells with fluorescently tagged GIP protein. Fluorescent microscopy revealed clear and distinct binding by GIP on the surface of MCF-7 cells. Therefore, these results provide conclusive evidence supporting the existence of physical interaction at the plasma membrane between GIP and estrogen dependent human breast cancer cells. One possible mechanism being explored for this interaction is the presence of a GIP-specific cell surface receptor.
Michael Popowich
Characterization of Quantum Dots as FRET Donors in Molecular Beacons
In an effort to develop new ways to detect gene expression a molecular beacon based assay was created. The molecular beacons utilized quantum dots as their FRET donors while using hexachlorofluorescein as the FRET acceptor. Steady state fluorescence and fluorescence lifetime measurements that were taken indicate that FRET does occur in the molecular beacons when a complementary sequence is not present. However the ability to reverse FRET when a complementary DNA sequence was added proved difficult and results were not clear. In an effort to get better reversal of FRET EcoRV was used to cut the Hex away from the QD when complementary DNA sequence was added. This improved the efficiency of FRET reversal.
Honorary Members:
- Charelle Carter- Advisor: Prof. Barbara Boyer, Biology
- Zohny Zohny- Advisor: Prof. Rob Olberg, Biology
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