Senior Project Advisor
John Antos
Document Type
Project
Publication Date
Spring 2024
Keywords
Sortase A, protein engineering, LC-MS, AlphaFold2
Abstract
Sortase A (SrtA) is a powerful protein engineering tool for the generation of site-specific unnatural polypeptide derivatives. We identified the N-terminal substrate element “DRIKE” as a motif which notably accelerates sortase-mediated ligation (SML) reactions in a variety of contexts defined by variables including nucleophile identity, SrtA identity, and N-terminal substrate domain type. Systematic DRIKE mutagenesis and AlphaFold2 models support a model in which the DRIKE P7 Ile makes a favorable hydrophobic contact with the SrtA enzyme surface, stabilizing enzyme-substrate binding. Similar sequence elements are present in natural SrtA substrate proteins, and these contexts also generally correlate with accelerated ligation relative to a pentaglycine control substrate. These findings implicate future high-throughput methods to discover optimal N-terminal SML substrates.
Department
Chemistry
Recommended Citation
Noah Goodwin-Rice, Noah, "Accelerating Sortase-Mediated Ligation Reactions with N-terminal Substrate Elements" (2024). WWU Honors College Senior Projects. 867.
https://cedar.wwu.edu/wwu_honors/867
Type
Text
Rights
Copying of this document in whole or in part is allowable only for scholarly purposes. It is understood, however, that any copying or publication of this document for commercial purposes, or for financial gain, shall not be allowed without the author’s written permission.
Language
English
Format
application/pdf