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.
Organisations
- MRC National Inst for Medical Research, United Kingdom (Lead Research Organisation)
- University of the Balearic Islands (Collaboration)
- Italian Institute of Technology (Istituto Italiano di Tecnologia IIT) (Collaboration)
- French National Institute of Agricultural Research (Collaboration)
- University of Naples (Collaboration)
- University of York, United Kingdom (Collaboration)
- Free University of Brussels (ULB), Belgium (Collaboration)
- PA Consulting Group (Collaboration)
People |
ORCID iD |
Annalisa Pastore (Principal Investigator) |
Publications

Pandolfo M
(2009)
The pathogenesis of Friedreich ataxia and the structure and function of frataxin
in Journal of Neurology

Sjekloca L
(2009)
A study of the ultrastructure of fragile-X-related proteins.
in The Biochemical journal

De Chiara C
(2009)
Phosphorylation of S776 and 14-3-3 binding modulate ataxin-1 interaction with splicing factors.
in PloS one

Sanfelice D
(2009)
Cold denaturation and aggregation: a comparative NMR study of titin I28 in bulk and in a confined environment.
in Journal of the American Chemical Society

Nicastro G
(2009)
Josephin domain of ataxin-3 contains two distinct ubiquitin-binding sites.
in Biopolymers

Zen A
(2009)
Using dynamics-based comparisons to predict nucleic acid binding sites in proteins: an application to OB-fold domains.
in Bioinformatics (Oxford, England)

Prischi F
(2009)
The N-terminus of mature human frataxin is intrinsically unfolded.
in The FEBS journal

De Rocco D
(2009)
MYH9 related disease: a novel missense Ala95Asp mutation of the MYH9 gene.
in Platelets

Adinolfi S
(2009)
Bacterial frataxin CyaY is the gatekeeper of iron-sulfur cluster formation catalyzed by IscS.
in Nature structural & molecular biology

Temussi PA
(2009)
Sweet, bitter and umami receptors: a complex relationship.
in Trends in biochemical sciences

Ivanova EV
(2008)
[Interface of the interaction of the middle domain of human translation termination factor eRF1 with eukaryotic ribosomes].
in Molekuliarnaia biologiia

De Chiara C
(2008)
Structural bases for recognition of Anp32/LANP proteins.
in The FEBS journal

Turjanski A
(2008)
NMR and molecular dynamics studies of the interaction of melatonin with calmodulin
in Protein Science

Rossetti G
(2008)
Structural properties of polyglutamine aggregates investigated via molecular dynamics simulations.
in The journal of physical chemistry. B

Riess O
(2008)
SCA3: neurological features, pathogenesis and animal models.
in Cerebellum (London, England)

Ehrnhoefer DE
(2008)
EGCG redirects amyloidogenic polypeptides into unstructured, off-pathway oligomers.
in Nature structural & molecular biology

Martin S
(2008)
Cold Denaturation of Yeast Frataxin Offers the Clue to Understand the Effect of Alcohols on Protein Stability
in Journal of the American Chemical Society

Correia AR
(2008)
Dynamics, stability and iron-binding activity of frataxin clinical mutants.
in The FEBS journal

Pastore A
(2007)
Peptides and proteins in a confined environment: NMR spectra at natural isotopic abundance.
in Journal of peptide science : an official publication of the European Peptide Society

Mantsyzov AB
(2007)
NMR assignments of the C-terminal domain of human polypeptide release factor eRF1.
in Biomolecular NMR assignments

De Chiara C
(2007)
NMR assignment of the leucine-rich repeat domain of LANP/Anp32a.
in Journal of biomolecular NMR

Pastore A
(2007)
Unbiased cold denaturation: low- and high-temperature unfolding of yeast frataxin under physiological conditions.
in Journal of the American Chemical Society

De Filippis V
(2007)
o-Nitrotyrosine and p-iodophenylalanine as spectroscopic probes for structural characterization of SH3 complexes.
in Protein science : a publication of the Protein Society

Tancredi T
(2007)
Conformation-activity relationship of neuropeptide S and some structural mutants: helicity affects their interaction with the receptor.
in Journal of medicinal chemistry

Pastore C
(2007)
Understanding the binding properties of an unusual metal-binding protein--a study of bacterial frataxin.
in The FEBS journal

D'ursi A
(2007)
Conformation-activity relationship of peptide T and new pseudocyclic hexapeptide analogs.
in Journal of peptide science : an official publication of the European Peptide Society


Pastore A
(2007)
A structural overview of the vertebrate prion proteins.
in Prion

Foury F
(2007)
Acidic residues of yeast frataxin have an essential role in Fe-S cluster assembly.
in EMBO reports

Klein FA
(2007)
Pathogenic and non-pathogenic polyglutamine tracts have similar structural properties: towards a length-dependent toxicity gradient.
in Journal of molecular biology

Goold R
(2007)
Down-regulation of the dopamine receptor D2 in mice lacking ataxin 1.
in Human molecular genetics

De Nicola G
(2007)
The structure of Lethocerus troponin C: insights into the mechanism of stretch activation in muscles.
in Structure (London, England : 1993)

Nicastro G
(2006)
Structure validation of the Josephin domain of ataxin-3: conclusive evidence for an open conformation.
in Journal of biomolecular NMR

Musi V
(2006)
New approaches to high-throughput structure characterization of SH3 complexes: the example of Myosin-3 and Myosin-5 SH3 domains from S. cerevisiae.
in Protein science : a publication of the Protein Society

Temussi PA
(2006)
Natural sweet macromolecules: how sweet proteins work.
in Cellular and molecular life sciences : CMLS

Bal NC
(2006)
NMR assignment of peptidyl-tRNA hydrolase from Mycobacterium tuberculosis H37Rv.
in Journal of biomolecular NMR

Polshakov VI
(2006)
Effects of co-operative ligand binding on protein amide NH hydrogen exchange.
in Journal of molecular biology

De Simone A
(2006)
Toward the understanding of MNEI sweetness from hydration map surfaces.
in Biophysical journal

Ramos A
(2006)
The structure of the N-terminal domain of the fragile X mental retardation protein: a platform for protein-protein interaction.
in Structure (London, England : 1993)

Correia AR
(2006)
Conformational stability of human frataxin and effect of Friedreich's ataxia-related mutations on protein folding.
in The Biochemical journal

Pastore C
(2006)
YfhJ, a molecular adaptor in iron-sulfur cluster formation or a frataxin-like protein?
in Structure (London, England : 1993)

Menon RP
(2006)
Expansion of amino acid homo-sequences in proteins: insights into the role of amino acid homo-polymers and of the protein context in aggregation.
in Cellular and molecular life sciences : CMLS

Esposito V
(2006)
The importance of electrostatic potential in the interaction of sweet proteins with the sweet taste receptor.
in Journal of molecular biology

Menon RP
(2006)
Hammerhead ribozyme-mediated silencing of the mutant fibrillin-1 of tight skin mouse: insight into the functional role of mutant fibrillin-1.
in Experimental cell research

Ivanova EV
(2006)
NMR assignments of the middle domain of human polypeptide release factor eRF1.
in Journal of biomolecular NMR

Fiorin G
(2006)
Using metadynamics to understand the mechanism of calmodulin/target recognition at atomic detail.
in Biophysical journal

Temussi P
(2006)
The history of sweet taste: not exactly a piece of cake.
in Journal of molecular recognition : JMR

Bulone D
(2006)
The interplay between PolyQ and protein context delays aggregation by forming a reservoir of protofibrils.
in PloS one

Thomas D
(2005)
WHEATSHEAF: an algorithm to average protein structure ensembles.
in Acta crystallographica. Section D, Biological crystallography

Fiorin G
(2005)
Unwinding the helical linker of calcium-loaded calmodulin: a molecular dynamics study.
in Proteins
Description | Co-chairperson in the annual grant evaluation for the Portuguese Science Ministry |
Geographic Reach | National |
Policy Influence Type | Participation in a guidance/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 a guidance/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 | Contribution to a national consultation/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 a guidance/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 a guidance/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 a guidance/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 a guidance/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) |
Funding ID | 242193 |
Organisation | European Commission |
Sector | Public |
Country | European Union (EU) |
Start | 05/2010 |
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 | 05/2015 |
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 | 08/2015 |
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 | Academic/University |
Country | United Kingdom |
Start | 09/2011 |
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 | 08/2015 |
End | 08/2018 |
Description | Newton Fellowship |
Amount | £96,000 (GBP) |
Organisation | The Royal Society |
Sector | Charity/Non Profit |
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) |
Funding ID | 603191 |
Organisation | European Commission |
Department | Seventh Framework Programme (FP7) |
Sector | Public |
Country | European Union (EU) |
Start | 03/2014 |
End | 03/2017 |
Description | RS-International Joint Project |
Amount | £12,000 (GBP) |
Organisation | The Royal Society |
Sector | Charity/Non Profit |
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 | 07/2011 |
End | 12/2011 |
Description | Understanding the aggregation properties of TDP43 in FTD |
Amount | £5,000,000 (GBP) |
Organisation | UK Dementia Research Institute |
Sector | Charity/Non Profit |
Country | United Kingdom |
Start | 08/2017 |
End | 08/2021 |
Description | University of Michigan (NIH) |
Amount | £39,775 (GBP) |
Organisation | National Institutes of Health (NIH) |
Sector | Public |
Country | United States |
Start | 05/2010 |
End | 05/2011 |
Description | University of Sussex |
Amount | £159,726 (GBP) |
Organisation | University of Sussex |
Sector | Academic/University |
Country | United Kingdom |
Start | 05/2008 |
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 | Rational design of RNA aptamers |
Organisation | Italian Institute of Technology (Istituto Italiano di Tecnologia IIT) |
Department | Neuroscience and Brain Technologies IIT |
Country | Italy |
Sector | Academic/University |
PI Contribution | We are developing RNA aptamers against TDP-43 |
Collaborator Contribution | The partner Gian Gaetnao Tartaglia is the author of the software used for the prediction. |
Impact | We have held together a grant from MNDA and published four papers together. We are now in the process of sending off a manuscript and a grant application to ADDF. |
Start Year | 2016 |
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 misfolding diseases: polyQ and prion diseases |
Organisation | French National Institute of Agricultural Research |
Country | France |
Sector | Academic/University |
PI Contribution | NMR spectrometr time, know-how on NMR structural determination |
Collaborator Contribution | know-how on light scatterins, calorimetry and kinetics. |
Impact | Two publications in prreparation |
Start Year | 2008 |
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 p 6 ALLERGEN AND THEIR USE |
Description | The invention relates to a hypoallergenic immunogenic molecule derived from the Phl p 6 allergen, wherein the Phl p 6 molecule has an N-terminal and/or C-terminal deletion which makes the molecule at least substantially lack IgE binding capacity. The invention also relates to a hypoallergenic immunogenic combination of molecules derived from the Phl p 6 allergen, comprising: (i) a Phl p 6 molecule having an N-terminal deletion which makes the molecule at least substantially lack IgE binding capacity, and (ii) a Phl p 6 molecule having a C-terminal deletion which makes the molecule at least substantially lack IgE binding capacity, which two molecules together encompass the complete sequence of Phl p 6. The invention further relates to the use of the hypoallergenic immunogenic molecule or molecule mixture in hyposensitization and diagnosis. |
IP Reference | WO0130816 |
Protection | Patent granted |
Year Protection Granted | 2001 |
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 | Trade Mark |
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 |