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Lsm proteins as ring tectons

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thesis
posted on 28.03.2022, 12:31 authored by Dominic Logel
Bionanotechnology is beginning to exploit naturally occurring proteins as building blocks for creating nanoscale materials and complex devices. One example of interest is the Lsm family of proteins, which spontaneously assemble in vivo either as homomeric (archaea or prokarya), or heteromeric (eukarya) oligomers comprising six or seven Lsm protomers which play a role in RNA metabolism. At Macquarie, several Lsm proteins have beeen reengineered to form novel artificial RNA-binding rings. In this thesis I explore biophysical characteristics of new versions of yeast-derived tectons: utilisng polyproteins Lsm[1+4] and Lsm[5+6]. For the first time, the Lsm[5+6]₄ tecton has been cleanly produced, and its stability evaluated, whilst pure Lsm[1+4] proved to be difficult to obtain. This thesis also describes the first steps towards a crystal structure of a ring assembly formed by R65PLsmα. This mutant has been proposed as a variant of the heptameric Lsmα₇, possibly comprising a hexameric organisation. I successfully generated many crystal forms, which led on to provide diffraction of good quality. Results from structure solving allowed definition of the correct number of components, which was not able to be determined from biophysical solution methods.

History

Table of Contents

Chapter 1. Form and architecture of Lsm proteins -- Chapter 2. Materials & methods -- Chapter 3. Characterisation of Lsm polyproteins as potential tectons -- Chapter 4. Crystallisation of an engineered Lsmα tecton -- Chapter 5. Conclusion.

Notes

Bibliography: pages a-e Empirical thesis.

Awarding Institution

Macquarie University

Degree Type

Thesis MRes

Degree

MRes, Macquarie University, Faculty of Science and Engineering, Department of Chemistry and Biomolecular Sciences

Department, Centre or School

Department of Chemistry and Biomolecular Sciences

Year of Award

2015

Principal Supervisor

Bridget Mabbutt

Rights

Copyright Dominic Logel 2015. Copyright disclaimer: http://www.copyright.mq.edu.au

Language

English

Extent

1 online resource (v, 45, a-e pages) illustrations (some colour)

Former Identifiers

mq:44909 http://hdl.handle.net/1959.14/1073155