Biomedical Engineering and Bioengineering Commons^™

Mutational Analysis Of Latency Associated Peptide For Therapeutic Application, Johnny Will Stephens

USF Tampa Graduate Theses and Dissertations

TGF-β is an important protein for regulation of the immune system, and has been linked to promotion of tumor progression and cancer growth. Inhibiting TGF-β has been shown to be an effective therapeutic technique for fighting multiple cancer types. Engineering TGF-β’s natural inhibitor, Latency Associated Peptide (LAP), to improve its biophysical properties has potential to increase therapeutic efficacy.

Rosetta and FoldX can be used in concert to engineer stabilizing mutations of proteins. In accordance with this, PyRosetta and FoldX were used to predict stabilizing mutations of LAP. The most stabilizing mutations from each program were combined with mutually predicted mutations. …

Engineering A Fluorescent Protease Sensor For In Vivo Protein Detection, Thomas C. Kinard

Honors Scholar Theses

This report details the results of an ongoing project to engineer a mutant form of Red Fluorescent Protein (RFP) variant mCherry that acts as a real-time in vivo protease sensor. The sought-after mutant only becomes fluorescent when exposed to Tobacco Etch Virus (TEV) Protease, this system’s model protease. This will be accomplished via the insertion of the TEV Protease Recognition Site (TEV-PRS) in such a position that, before cleavage, will prevent the protein from folding to fluorescent conformation, but upon cleavage, will allow for fluorescent conformation to occur. The cylindrical structure of the protein, composed of beta-pleated sheets, contains “loops” …