UVM Theses and Dissertations
Format:
Print
Author:
Banerjee, Sunandan
Dept./Program:
Chemistry
Year:
2012
Degree:
PhD
Abstract:
The basic premise of this dissertation is to develop a methodology for screening optimally stable peptide secondary structures on solid-phase synthesis resins based on hydrophobic core packing. Initial studies on assembling metal-assisted 3-helix bundle proteins on these resins indicated the need to chemically modify the resin surface for exchange of peptide modules between solution and the resin surface. This led to a study of behavior of different peptides and proteins on various resin surfaces displaying novel functionalities to minimize protein resistance. Ribose-derivatized resin surfaces were discovered as novel solution to the problem of protein adsorption on resin surfaces.
To obtain proof-of-concept for assembling stable protein motifs on resins, the protein motif of choice is coiled-coils, found widely amongst a class of proteins called transcription factors which control DNA transcription and hence protein synthesis in cells. The experimental design involves synthesizing libraries of coiled-coil peptides in solution and on resin beads. Allowing these libraries to screen for optimally stable partnerships enables the sequestration of selective non-covalent complexes on the polymer beads, which are then identified through mass spectrometry. The results obtained indicate that the best possible partnerships are indeed between hydrophobic residues thereby supporting the idea that stable protein architectures can be assembled on solid-phase synthesis resins based upon hydrophobic core packing of individual peptide motifs.
To obtain proof-of-concept for assembling stable protein motifs on resins, the protein motif of choice is coiled-coils, found widely amongst a class of proteins called transcription factors which control DNA transcription and hence protein synthesis in cells. The experimental design involves synthesizing libraries of coiled-coil peptides in solution and on resin beads. Allowing these libraries to screen for optimally stable partnerships enables the sequestration of selective non-covalent complexes on the polymer beads, which are then identified through mass spectrometry. The results obtained indicate that the best possible partnerships are indeed between hydrophobic residues thereby supporting the idea that stable protein architectures can be assembled on solid-phase synthesis resins based upon hydrophobic core packing of individual peptide motifs.