Structural studies of proteins involved in neurodegenerative and muscular diseases

Lead Research Organisation: MRC National Inst for Medical Research

Abstract

The accomplishment of the human genome together with the fast progresses made in gene mapping technology have opened for the first time the possibility of approaching hereditary disorders at a molecular level. We approach the study of human diseases by understanding their structure/function relationship. The role of the molecular structure in modern biology and medicine may be directly appreciated if we consider that structure determination of the DNA, the molecule which contains the whole genetic information, unveiled the secret of life. Our laboratory focuses on determining the 3D structure of proteins with specific interest for proteins involved in neurodegenerative diseases such as the Huntington Chorea, Friedreich ataxia, and Fragile X syndrome and muscular pathologies such as different types of cardio-myopathies. Despite the different symptoms, these diseases, all incurable, share the fact that in affected patients, the proteins encoded by the corresponding genes are produced in altered forms, or in smaller quantity down to their complete absence. By determining the shape of the proteins involved we can then understand the role of these mutations on the protein functions and ultimately design new therapeutical strategies. We use a combination of biochemical and biophysical techniques which help us to reconstruct the cellular role of the proteins of interest and characterise their structure/function relationship

Technical Summary

We focus on neurodegeneration and muscular pathologies.||A molecular approach to human genetic diseases More than 12 human genetic diseases have been associated with expansions of nucleotide repeats in their corresponding genes, all identified in the last 10 years. These diseases, currently incurable, are interesting both for their impact in the human population and for the unusual pattern of genetic behaviour. They are grouped into two sub-families according to whether the expansion occurs in coding or non-coding regions. In our group we have started an integrated approach which makes use of different biochemical and biophysical techniques to study the structure and function of the proteins encoded by several genes of the trinucleotide expansion family. Amongst them are FMRP, the protein responsible for fragile x and frataxin, the protein involved for Friedreichs ataxia. We want to understand the cellular role of these proteins and describe the mechanism of molecular recognition with their binding partners. We have also recently setup a systems biology project which aims at understanding the process of iron-sulfur cluster formation, in which frataxin is involved.||How muscles are assembled and regulated Understanding the molecular bases of muscle contraction has been one of the main research topics in biology for the last decades. Besides actin and myosin, which are the main players in muscle contraction, many other proteins have an essential role in determining muscle assembly and regulation. Several interactions have been described recently which are essential to anchor the muscle filaments to the modular proteins, such as titin, nebulin, and alpha-actinin. Dysfunctions in the interactions cause different types of cardio-myopathies and other muscle diseases. We are currently working to determine the structure of a number of protein complexes. This information will eventually allow us to reconstruct the muscle ultrastructure.

Publications

10 25 50
 
Description Co-chairperson in the annual grant evaluation for the Portuguese Science Ministry
Geographic Reach National 
Policy Influence Type Participation in advisory committee
Impact We advised the government on the distribution of National grants
 
Description Consultant for the evaluation of National Grants in Italy (MURST, MIUR, PRIN), Belgium, France (Pasteur Institute, BLANC), UK (Wellcome, MRC, BBSRC), Swiss National Science Foundation, Portugal and Spain
Geographic Reach Asia 
Policy Influence Type Participation in advisory committee
Impact I refereed National Grants
 
Description Member of an International panel for the assessment of Italian Scientists organized by the Italian Ministry
Geographic Reach National 
Policy Influence Type Gave evidence to a government review
Impact We advised the Government on the distribution of National Grants
 
Description Panel member in a quinquennial evaluation of the Forth (Eraklion) and Fleming Institutes (Athens)
Geographic Reach National 
Policy Influence Type Participation in advisory committee
Impact We advised the Greek government on these Institutions.
 
Description Panel member of a DFG assessment to decide on applications for high field (1.2 GHz) spectrometers in Germany
Geographic Reach National 
Policy Influence Type Participation in advisory committee
Impact We advised the Government on the purchase of 1.2GHz NMR spectrometers in Munich, Frankfurt and Gottingen
 
Description Panel member of the Roadmap Working Group of Experts for Biological and Medical Sciences (EU grants)
Geographic Reach Asia 
Policy Influence Type Participation in advisory committee
Impact We advised EU on the creation of new Infrastructures
 
Description Panel member of the Scientific Advisory Board of Biotechnology Institute in Helsinki, Finland
Geographic Reach Asia 
Policy Influence Type Participation in advisory committee
 
Description Ataxia (UCL)
Amount £36,000 (GBP)
Organisation Ataxia UK 
Sector Charity/Non Profit
Country United Kingdom
Start 02/2009 
End 01/2012
 
Description BC-British-Italian Partnership Programme-2006
Amount £1,040 (GBP)
Organisation British Council 
Sector Charity/Non Profit
Country United Kingdom
Start  
 
Description BC-Travel Grant
Amount £1,000 (GBP)
Organisation British Council 
Sector Charity/Non Profit
Country United Kingdom
Start 01/2006 
End 12/2007
 
Description BC-Travel Grant-2006
Amount £500 (GBP)
Organisation British Council 
Sector Charity/Non Profit
Country United Kingdom
Start 01/2006 
End 12/2008
 
Description EFACTS 2010-2014
Amount £350,000 (GBP)
Organisation European Commission 
Sector Public
Country European Union (EU)
Start 06/2010 
End 05/2014
 
Description FARA (Frataxin Research)
Amount £28,291 (GBP)
Organisation Ataxia UK 
Sector Charity/Non Profit
Country United Kingdom
Start 01/2006 
End 12/2008
 
Description Fellowship
Amount £18,000 (GBP)
Organisation Kids company 
Sector Charity/Non Profit
Country United Kingdom
Start 06/2015 
End 08/2015
 
Description Grant
Amount $35,000 (USD)
Organisation National Ataxia Foundation 
Sector Charity/Non Profit
Country United States
Start 03/2015 
End 08/2015
 
Description Grant
Amount £245,000 (GBP)
Organisation Biotechnology and Biological Sciences Research Council (BBSRC) 
Sector Public
Country United Kingdom
Start 09/2015 
End 08/2018
 
Description MRC Research Grant
Amount £1,074,682 (GBP)
Organisation Medical Research Council (MRC) 
Department MRC National Institute for Medical Research (NIMR)
Sector Public
Country United Kingdom
Start 10/2011 
End 04/2017
 
Description Motor Neuro Disorder Association
Amount £221,595 (GBP)
Organisation Motor Neurone Disease Association (MND) 
Sector Charity/Non Profit
Country United Kingdom
Start 09/2015 
End 08/2018
 
Description Newton Fellowship
Amount £96,000 (GBP)
Organisation The Royal Society 
Sector Academic/University
Country United Kingdom
Start 11/2015 
End 10/2017
 
Description Novexin-Research Grant2006
Amount £500 (GBP)
Organisation Expedeon Protein Solutions (formerly Novexin Ltd) 
Sector Private
Country United Kingdom
Start 01/2006 
End 12/2007
 
Description Paincage
Amount € 400,000 (EUR)
Organisation European Commission 
Department Seventh Framework Programme (FP7)
Sector Public
Country European Union (EU)
Start 04/2014 
End 03/2017
 
Description RS-International Joint Project
Amount £12,000 (GBP)
Organisation The Royal Society 
Sector Academic/University
Country United Kingdom
Start 01/2006 
End 12/2007
 
Description Studies of the effects of glycation on amylin aggregation
Amount £15,000 (GBP)
Organisation Diabetes UK 
Sector Charity/Non Profit
Country United Kingdom
Start 08/2011 
End 12/2011
 
Description Understanding the aggregation properties of TDP43 in FTD
Amount £5,000,000 (GBP)
Organisation UK Dementia Research Institute 
Sector Academic/University
Country United Kingdom
Start 09/2017 
End 08/2021
 
Description University of Michigan (NIH)
Amount £39,775 (GBP)
Organisation National Institutes of Health (NIH) 
Sector Public
Country United States
Start 06/2010 
End 05/2011
 
Description University of Sussex
Amount £159,726 (GBP)
Organisation University of Sussex 
Sector Academic/University
Country United Kingdom
Start 06/2008 
End 05/2011
 
Title Production of mylated peptides 
Description We developed a robust method to glycate peptides 
Type Of Material Biological samples 
Provided To Others? No  
Impact This allows the facile production of peptides with this post-translational modification 
 
Title Production of post-translational modification in proteins 
Description We designed a strategy to ubiquitinate ataxin-3 
Type Of Material Biological samples 
Year Produced 2013 
Provided To Others? Yes  
Impact We have provided a new methodology to produce suitable amounts of mono-ubiquitinated ataxin-3 
 
Title a new model for the causes of Friedreich's ataxia 
Description We identified the function of frataxin and provided a model for how its absence causes the disease 
Type Of Material Model of mechanisms or symptoms - in vitro 
Year Produced 2009 
Provided To Others? Yes  
Impact We provided information on how to mutate proteins for validating our model. 
 
Description Protein stability and cold denaturation 
Organisation University of Naples
Country Italy 
Sector Academic/University 
PI Contribution We have provided spectrometer access, protein material, know-how on NMR measurements.
Collaborator Contribution Protein stability, crowding and confinement.
Impact Five pubblications (163) Sanfelice, D., Tancredi, T., Politou, A., Pastore, A. And Temussi, P. (2009) Cold denaturation and aggregation: a comparative NMR study for titin I28 in bulk and in a confinded environment. JACS 131, 11662-11663. (157) Esposito, V., Pastore, A., Martin, S., de los Rios, P., Temussi, P.A. (2008) Cold denaturation of yeast frataxin offers the clue to understand the effect of alcohols on protein stability.JACS 130, 9963-9970. (152) Pastore, C., Franzese, M., Sica, F., Temussi, P. and Pastore, A. (2007) Understanding the binding properties of an unusual metal binding protein: a study of bacterial frataxin. Febs J. 274, 4199-4210 (150) Pastore, A., Martin, S.R, Politou, A., Kondapalli, K.C., Stemmler, T., Temussi, P. A. (2007) Unbiased cold denaturation: low and high temperature unfolding of Yeast frataxin under physiological conditions. JACS 129; 5374-5375. (147) Pastore, A., Salvadori, S., Temussi, P.A. (2007) Peptides and proteins in a confined environment: NMR spectra at natural isotopic abundance. J. Pept. Sci. 15, 795-807.
Start Year 2006
 
Description Studies of how glycation influences cell toxicity 
Organisation University of Naples
Country Italy 
Sector Academic/University 
PI Contribution We have a long term experience in understanding the factors that influence aggregation propensity
Collaborator Contribution The Italian group is currently working on the effects of glycation on toxicity. we therefore send them glycated amylin and Abeta to test for roxicity.
Impact We have complemented our studies on the structural and aggregation effects of glycation.
Start Year 2014
 
Description Studies of how glycation influences the aggregation propensityof polypeptides 
Organisation University of the Balearic Islands
Country Spain 
Sector Academic/University 
PI Contribution We have a long term experience in production of recombinant proteins and peptides. We have helped their group to set up a lab for this purpose and sent there one of our postdoc for 3 months to instruct the researchers there.
Collaborator Contribution The group there has been working for several years on glycation. Their contribution has been invaluable for starting our research for the DiabetesUK grant. We are also constantly exchange expertise, know-how, protocols and materials.
Impact We have been able to smoothly start the work planned for this grant. We have produced preliminary data which were crucial to obtain the current grant (started only 10 months ago).
Start Year 2011
 
Description Studies of the frataxin function 
Organisation Free University of Brussels
Country Belgium 
Sector Academic/University 
PI Contribution We have developed an assay which has allowed us to elucidate the function of frataxin
Collaborator Contribution co-authored publications and in vivo experiments to test our functional hypotheses
Impact As a result of this collaboration we are now part of an EU grant awarded for 2009-2012. Pandolfo, M., Pastore, A. (2008) The pathogenesis of Friedreich ataxia and the structure and function of frataxinJ. of Neurol. 256 Suppl 1:9-17
Start Year 2010
 
Description The mechanism of protein aggregation 
Organisation Moscow State University
Department Biochemistry Department
Country Russian Federation 
Sector Academic/University 
PI Contribution We have been collaborating on the effects of metals on the aggregation of the Abeta peptides.
Collaborator Contribution They produced the peptide and analysed the data collected in London
Impact We have been accepted a paper in Scientific reports.
 
Description Understanding the structural bases of stretch activation in muscle 
Organisation University of York
Country United Kingdom 
Sector Academic/University 
PI Contribution Biophysical studies, structural know-how.
Collaborator Contribution Material, know-how and exhange of personnel
Impact (117) De Nicola, G., Biekofsky, R., Kelly, G., Agianian, B., Bullard, B., Pastore, A. (2004) Assignment of the 1H, 13C, and 15N resonances of holo isoform 4 of Lethocerus indicus Troponin C. J. of Biomol. NMR, 29, 461-462. (151) DeNicola, G., Burkart, C., Qiu, F., Agianian, B., Labeit, S., Martin, S., Bullard, B., Pastore, A. (2007) The structure of Lethocerus troponin C: New insights into the mechanism of stretch activation in muscles. Structure 15, 813-824.
Start Year 2006
 
Title Non-anaphylactic forms of grass pollen Ph1 p6 allergen and their use 
Description Design of a Non-anaphylactic forms of grass pollen Ph1 p6 allergen and their use 
IP Reference WO2001030816 
Protection Patent granted
Year Protection Granted
Licensed Yes
Impact This provides the bases for drug development against grass pollen allergies.
 
Title the function of frataxin 
Description We have identified the function of frataxin and its role in friedreich's ataxia 
IP Reference  
Protection Protection not required
Year Protection Granted 2009
Licensed No
Impact Our research has completely revolutionaised the field of FRDA research
 
Description Press release 
Form Of Engagement Activity A press release, press conference or response to a media enquiry/interview
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Media (as a channel to the public)
Results and Impact Press release for our work on Friedreich's ataxia

The promotion of reserach on this disease. The establishment of a EU network to support the research further.
Year(s) Of Engagement Activity 2009
 
Description Publication of a science dissemination in Frontiers for young minds 
Form Of Engagement Activity A magazine, newsletter or online publication
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Schools
Results and Impact I published an article in frontiers for Young Minds which is directed to children in the range of 8 to 16 years old. This initiative is meant to disseminate research among young people.
Year(s) Of Engagement Activity 2017