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 College Senior Projects. 398.
Subjects - Topical (LCSH)
student projects; term papers
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