Senior Project Advisor
Sortase A, peptide, unstructured loop, cleavage activity, trapping peptide, LPXTG, synthesis, unnatural peptide, crystallization
Sortase A is a powerful protein engineering tool that cleaves proteins and attaches them to an acyl acceptor of choice. However, the most active wild type variants of sortase A only show high activity at a limited number of cleavage motifs, and so work is underway to create a variant of sortase A that shows high activity at a greater variety of cleavage sites. Additionally, current studies attempting to optimize the enzyme require a way to stabilize an unstructured loop for the crystallization, in order to collect X-ray diffraction structural data. Here, preliminary results from the design of an “loop-swapped” sortase A enzyme variants are discussed, as well as the synthesis of two different thiol-containing “trapping” peptides intended to freeze sortase A in its bound conformation via a covalent disulfide bond. The first peptide is a previously described unnatural peptide, and was discontinued before completion, and the second peptide is a natural peptide which was completed and purified. Future work will involve the co-crystallization of this peptide with “loop-swapped” sortase A variants.
Johnston, Katherine, "Sorting Out Sortases: Designing a Better Enzyme and Synthesizing Trapping Peptides Towards Capturing a Bound-State Structure" (2020). WWU Honors Program Senior Projects. 398.
student projects; term papers
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.