Developing a Hydrogen/Deuterium Exchange Mass Spectrometry (HDX-MS) Platform to Study the Structure and Assembly of Bacterial Nanocompartments.

Lead Research Organisation: University of Edinburgh
Department Name: Sch of Chemistry

Abstract

Bacterial nanocompartments (BNCs) are large protein assemblies comprising a hollow icosahedral multimeric protein cage which can sequester a range of 'cargo' protein (often enzymes) inside. This project will develop techniques in hydrogen/deuterium exchange mass spectrometry (HDX-MS) to study the structure, dynamics and assembly of BNCs. These mass spectrometry studies will complement existing structural biology studies using X-ray crystallography and electron microscopy. Understanding these fascinating systems will allow their future manipulation as potential platforms for biotechnology, synthetic biology, and drug delivery.
The student will work in a highly interdisciplinary team spanning the disciplines of chemistry, biology and engineering. The student will learn skills in protein mass spectrometry, recombinant protein production, biochemistry, and structural biology. Protein crystallography and electron microscopy will be performed in collaboration with Dr. Jon Marles-Wright, University of Newcastle (www.marles-wright-lab.org).

Publications

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Studentship Projects

Project Reference Relationship Related To Start End Student Name
EP/N509644/1 01/10/2016 30/09/2021
1940602 Studentship EP/N509644/1 01/09/2017 28/02/2021 Thomas Lambert
 
Description Ferritins are a large family of intracellular proteins involved in iron oxidation and storage. Produced by almost all living organisms Ferritins oxidize iron from Fe(II) to Fe(III) then store them within their spherical quaternary structure. Encapsulated ferritins (EncFtn) are a subfamily of ferritins that are sequestered into larger macrostructure Encapsulin capsids. It is thought that this interplay between the EncFtn and the Encapsulin can be repurposed into an effective drug delivery system. However, questions remain regarding the assembly pathways of both EncFtn and the larger Encapsulin-EncFtn complex. We have used HDX-MS to further elucidate how the conformation of the EncFtn dimer is essential for the formation of its higher order oligomeric states.
The award has resulted in the Confirmation of role the dimeric assemblies within Encapsulated ferritin have to the decameric assembly of Encapulated Ferritin via HDX-MS.
Exploitation Route The with automated platform built and programmed the departments HDX-MS capabilities have increased significantly. This platform can now be used as a basis for further research and or a starting point for new PhD student to continue the work on HDX-MS analysis on Encapsulin nano-compartments.
Sectors Manufacturing, including Industrial Biotechology,Pharmaceuticals and Medical Biotechnology