Chaperone-assisted protein folding in the bacterial periplasm

Lead Research Organisation: University of Leeds
Department Name: Astbury Centre


Molecular chaperones assist the folding of virtually all cellular proteins. Despite years of intensive study, the fundamental question of how chaperones facilitate protein folding remains unclear. Whilst molecular chaperones in the cytosol use the energy of ATP hydrolysis to control substrate binding, folding and release, how chaperones in the bacterial periplasm assist folding in the absence of ATP is unknown. Recent experiments have identified the substrates of the periplasmic chaperones Spy, Skp and SurA. Some of these chaperones assist folding of both water-soluble and membrane proteins, raising fascinating questions about how a chaperone is able to recognize its substrates, to release them in a controlled fashion and to assist folding. In this project the student will use biochemical and biophysical methods to determine how periplasmic molecular chaperones recognize their substrates, assist folding and, in tandem with each other and the Bam complex, create the folding pipeline required for bacterial growth and survival.


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Studentship Projects

Project Reference Relationship Related To Start End Student Name
BB/M011151/1 30/09/2015 29/09/2023
1646705 Studentship BB/M011151/1 30/09/2015 29/09/2019
Description Developed a new method for analysing the interactions between two proteins at high temporal and spatial resolution using cross-linking mass spectrometry.

Discovered the mechanism by which chaperones in bacteria prevent the aggregation of their outer membrane protein substrates.
Exploitation Route The new methodology that has been developed will be useful for researchers across the globe studying protein-protein interactions, the methodology has been published and the chemical reagents that were designed are now commercially available.
Sectors Chemicals,Pharmaceuticals and Medical Biotechnology

Title Creation of Tag-Transfer crosslinkers 
Description Developed new chemical crosslinking reagents for high resolution mapping of protein-protein interfaces in dynamic and static complexes. Developed method for using and analysing data generated by this method. 
Type Of Material Technology assay or reagent 
Year Produced 2018 
Provided To Others? Yes  
Impact Used in my research on outer membrane protein biogenesis. Used by other researchers studying mechanisms of amyloid formation / protein misfolding (Prof. Sheena Radford, University of Leeds). Used in the study of protein-protein interactions (Prof. Andy Wilson, University of Leeds). Reagents are commercially available for use by researchers across the globe from Redbrick Molecular Ltd. 
Description Central Laser Facility cryoSTORM of Outer Membrane Proteins 
Organisation Rutherford Appleton Laboratory
Department Central Laser Facility
Country United Kingdom 
Sector Academic/University 
PI Contribution Provided samples, intellectual input and an impactful research question to be applied to a newly developed / in development technique by the group of Prof Marisa Martin-Fernandez at the Central Laser Facility
Collaborator Contribution Provided access to resources and training in a newly developed technique, cryogenic STORM, for imaging bacterial outer membrane proteins at a nanometre scale. Allows us to visualise membrane organization with a better localisation precision than standard room temperature super-resolution microscopy techniques and allows fixation of samples in a frozen-hydrated (more physiological) state than chemical fixants.
Impact Poster presented at the 'Imagine: Imaging Life 2018' symposium at the University of Sheffield.
Start Year 2018