Skip navigation
Please use this identifier to cite or link to this item:
Title: Grafting Epitopes into a Disulfide-Constrained Tail of a Lasso Peptide
Authors: Chen, Cathy
Advisors: Link, A. James
Department: Chemical and Biological Engineering
Class Year: 2014
Abstract: Bioactive peptides are a promising direction for drug development. The specific goal of this study was to add a bioactive sequence to the C-terminus of the lasso peptide astexin-3 (Atx-3). Two sequences were tested, both containing the motif WEWEC. This motif inhibits activity of vascular endothelial growth factor, VEGF, an extracellular ligand produced in excess by cancer cells. The results of this study use mass spectroscopy to show the potential for adding a bioactive sequence to the C-terminus of Atx-3. The first sequence (WEWEC), called short-WEWEC, was successfully inserted into an existing Atx-3 vector and induced to produce varying levels of the full-length peptide. The lower temperature (16 degrees C) produced relatively higher signals of the full-length peptide at 3227 m/z and decreased C-terminus truncation. A longer induction time (48 hours) was needed for peptide production and for disulfide bond formation. The long-WEWEC sequence (IHVMWEWEC) was also successfully inserted into an existing Atx-3 vector. Similarly, a lower temperature (16 degrees C) and longer times of induction resulted in potential dimerization and decreased truncation of the full-length peptide. Although the full-length long-WEWEC peptide (Atx-3 with IHVMWEWEC addition to C-terminus) wasn’t observed, it appears the truncation intermediates favored three or four additional residues at the tail of the original Atx-3 peptide.
Extent: 53 pages
Type of Material: Princeton University Senior Theses
Language: en_US
Appears in Collections:Chemical and Biological Engineering, 1931-2016

Files in This Item:
File SizeFormat 
Chen_Cathy_CBE 14_Thesis Final.pdf1.69 MBAdobe PDF    Request a copy

Items in Dataspace are protected by copyright, with all rights reserved, unless otherwise indicated.