Strategies for Computational Protein Design with Application to the Development of a Biomolecular Tool-kit for Single Molecule Protein Sequencing

<p>One of the key properties of proteins is that they exhibit remarkable affinities and specificities for small-molecule and peptide binding partners. To improve the success rate of rational, computational protein design and widen the scope of potential applications, it is useful to define generalized strategies and automated methodology to improve and/or alter the affinity and specificity of interactions. I have implemented several strategies for engineering protein-small molecule interactions including: improvement of substrate accessibility, stabilization of the bound state, truncation and surface engineering, and transplantation of residue level, native (or native-like) interactions. Each strategy was applied to one or more model protein, and the resulting changes in affinity, specificity, and activity were characterized experimentally. Finally, we designed a biomolecular tool-kit, consisting of 17 engineered proteins for amino acid side-chain recognition and a single enzyme to catalyze the Edman degradation. We profiled the affinity and specificity of each protein, and implemented a computational framework that demonstrates its utility for amino acid calling in a single molecule protein sequencing assay.</p>

Name thesis:degree_name

Doctor of Philosophy (PhD)

Level thesis:degree_level

Dissertation

Discipline thesis:degree_discipline

Biology and Biomedical Sciences: Computational and Systems Biology

Year dc:date.available

2014

Author dc:creator

• Borgo, Benjamin

Contributors dc:contributor

• James J Havranek

• Protein design
• Protein engineering
• Protein sequencing
• Proteomics

Language dc:language

English (en)

DOI

https://doi.org/10.7936/K7610X9N

OAI identifier oai:identifier

oai:openscholarship.wustl.edu:etd-2221