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MRC Prion Unit
From fundamental research to prevention and cure
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Structural studies of prion proteins and their ligand interactions

A major programme in the Unit focuses on the central problem in understanding prions; what is the change in shape that distinguishes normal prion protein, PrPC, from its rogue form, PrPSc, and how does it come about? The team, led by Professor Tony Clarke, works mainly with the human prion protein itself - which is synthetically produced in the Unit in large quantities (using genetically engineered bacteria) in a specially-designed laboratory. A range of sophisticated equipment is used to study the structure, folding and dynamics of prion protein both in isolation and also with likely binding partners, in order to understand how it changes its shape.

These studies have yielded detailed information on the interaction between prion protein and a highly specific antibody which has been used successfully in the diagnosis of prion disease, and in experimental models has prevented the onset of disease symptoms following prion infection1. This has provided important clues as how prion protein molecules may interact with each other and how certain regions of prion protein may “seed” or start the disease process.

Structure of PrP bound to a therapeutic antibody

Structure of PrP bound to a therapeutic antibody. The complex between Prion protein and a highly-specific antibody as determined by x-ray crystallography. (A) PrP (green) with Antibody heavy and light chains (cyan and magenta, respectively). (B) Expanded view of the Prion Protein/Antibody  interface. PrP residues are labelled in black, those from the Antibody heavy and light chains in blue and magenta, respectively. Potential hydrogen bonds are shown as dashed lines.


We have also shown that the influence of a very strong and common genetic risk factor at position 129 in prion protein is not effected through a change in the shape or stability of the normal form of prion protein, PrPC. Instead, its effect is mediated through other alternatively shaped forms of the protein downstream in the disease process2,3. A number of these alternate “intermediate” states have been characterised by us, under conditions where they are stable, and also when they transiently-formed. These studies have highlighted regions of PrP which appear to direct the folding process, but which if perturbed can result in the formation of aberrantly-shaped forms which may facilitate formation of the disease-causing form of prion protein4,5.

Three-dimensional structure of human prion protein

Three-dimensional structure of human prion protein
, with the position ofthe common genetic risk factor highlighted in yellow.


Our work has also focused on the interactions that prion protein has with other molecules, for instance its role in copper metabolism6-9. A major aspect of this work is a collaboration between the Unit and GlaxoSmithKline (funded by the Department of Health) with the key aim of identifying compounds that will form the basis for anti-prion drugs. A simple test was developed that allowed the evaluation of over 1 million compounds for their ability to prevent the conversion of normal prion protein to the rogue form. This research programme is crucial to our efforts to produce effective drugs to block prions.

Conditions have been found that allow an engineered form of the prion protein to be converted into a form similar to that of the rogue PrPSc form10. This work has formed the basis for the production of the range of highly specific antibodies that have been used successfully in the diagnosis of prion disease {Beringue et al, 2004, Khalili-Shirazi, et al, 2005ab, 2007, Schimmel, et al, 2002}. This “synthetic” rogue prion forms large structures similar to those found in diseased brains, which has allowed high-resolution imaging of the type of structures that are formed in the prion disease process11. In addition, this “synthetic” prion analogue is a potent inhibitor of part of a key part of a cell which has been implicated in a number of neurodegenerative diseases12, providing further clues as to the prion disease mechanism. Work on determining the structure of this form of the protein is in progress at the unit13.

3D reconstruction of PrP fibrils

3D reconstruction of PrP fibrils formed from the engineered form of the prion protein

Accidental transmission of prion disease, via contaminated surgical instruments has occurred as prions resist conventional hospital sterilisation methods. In order to address this problem, we have developed a enzymatic method for the decontamination of infectious prion material adhered to surgical steel surfaces, which is inexpensive, non-corrosive to instruments, non-hazardous to staff and compatible with current equipment and procedures used in hospital sterilization units14. This successful translational development has now been licensed to the DuPont Corporation who are producing practical products to improve patient safety.


Peer reviewed articles:
2014

Prion neuropathology follows the accumulation of alternate prion protein isoforms after infective titre has peaked
Sandberg M, Al-Doujaily H, Sharps B, Wiggins De Oliveira M, Schmidt C, Richard-Londt A, Lyall S, Linehan J, Brandner S, Wadsworth J, Clarke A, Collinge J. Prion Nat Commun 2014

N-terminal domain of prionprotein directs its oligomeric association
Trevitt C, Hosszu L, Batchelor M, Panico S, Terry C, Nicoll A, Risse E, Taylor W, Sandberg M, Al-Doujaily H, Linehan J, Saibil H, Scott D, Collinge J, Waltho J, Clarke A. 
Journal of Biological Chemistry. 2014, Jul 29.Epub ahead of print. 

2011

Prion propagation and toxicity in vivo occur in two distinct mechanistic phases
Sandberg MK, Al-Doujaily H, Sharps B, Clarke AR, Collinge J.  Nature 2011; 470: 540-2.

Rapid cell-surface prion protein conversion revealed using a novel cell system
Goold R, Rabbanian S, Sutton L, Andre R, Arora P, Moonga J, Clarke AR, Schiavo G, Jat P, Collinge J, Tabrizi SJ. Nat Commun 2011; 2: 281.

Interaction between cellular prion protein and toxic Aβ oligomers can be therapeutically targeted at multiple sites
Freir DB, Nicoll AJ, Klyubin I, Panico S, Mc Donald JM, Risse E, Asante EA, Farrow -MA, Sessions RB, Saibil HR, Clarke AR, Rowan MJ, Walsh DM Collinge J. Nat Commun 2011; 2: 336

2010

PAW35 Anti-prion protein monoclonal antibodies at low doses effectively treat prion disease in mice without side-effects
Carswell C, Khalili-Shirazi A, Brandner S, Martins S, Drynda R, Collinge J, Mead S, Clarke A.
J Neurol Neurosurg Psychiatry 2010; 81: e33.

Pharmacological chaperone for the structured domain of human prion protein
Nicoll A, Trevitt C, Risse E, Quaterman E, Ibarra AA, Wright C, Jackson GS, Sessions R, Farrow M, Waltho J, Clarke A, Collinge J.  Proc Natl Acad Sci USA 2010; 107; 17610-5. 

The H187R mutation of the human prion protein induces conversion of recombinant prion protein to PrPSc-like form
Hosszu L, Tattum H, Jones S, Trevitt CR, Wells MA, Waltho JP, Collinge J, Jackson GS, Clarke AR.
Biochemistry 2010; 49: 8729-38.

2009

Highly sensitive, quantitative cell-based assay for prions adsorbed to solid surfaces
Edgeworth JA, Jackson GS, Clarke AR, Weissmann C, Collinge J.  Proc Natl Acad Sci USA 2009; 106: 3479-83.

Folding kinetics of the human prion protein probed by temperature jump.
Hart T, Hosszu LL, Trevitt CR, Jackson GS, Waltho JP, Collinge J, Clarke AR.
Proc Natl Acad Sci USA 2009; 106: 5651-6.

Conformational properties of beta -PrP
Hosszu LL, Trevitt CR, Jones S, Batchelor M, Scott DJ, Jackson GS, Collinge J, Waltho JP, Clarke AR.  J Biol Chem 2009; 284: 21981-90.

Crystal structure of human prion protein bound to a therapeutic antibody
Antonyuk SV, Trevitt CR, Strange RW, Jackson GS, Sangar D, Batchelor M, Cooper S, Fraser C, Jones S, Georgiou T, Khalili-Shirazi A, Clarke AR, Hasnain SS, Collinge J. Proc Natl Acad Sci USA 2009; 106: 2554-8.


Major external collaborations:

Professor Jonathon P Waltho,
Biomolecular NMR, University of Sheffield

Professor Helen Saibil,
Department of Crystallography, Birkbeck College and Institute of Structural Molecular Biology, University of London

Dr. Katherine McAuley,
Diamond Light Source, Harwell Campus, Oxfordshire

Professor Ian Collinson,
Department of Biochemistry, University of Bristol

Dr Richard Sessions,
Department of Biochemistry, University of Bristol

Professor Mervyn Miles,
Centre for Nanoscience and Quantum Information, University of Bristol

Professor Leo Brady,
Department of Biochemistry, University of Bristol 

Dr David Scott,
National Centre for Macromolecular Hydrodynamics, University of Nottingham

GlaxoSmithKline

MRC Prion News