Senior Project Advisor

Amacher, Jeaninie

Document Type

Project

Publication Date

Spring 2020

Keywords

Sortase A, peptide, unstructured loop, cleavage activity, trapping peptide, LPXTG, synthesis, unnatural peptide, crystallization

Abstract

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.

Department

Chemistry

Genre/Form

student projects; term papers

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

Included in

Chemistry Commons

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