PINT

Scientific supervisor: 
Description: 

Logo FEDER Région Sud

The INteractome Timone Platform (PINT) offers unique expertise in thermodynamic and kinetic analysis of molecular interactions.

PINT is equipped with microcalorimeters (ITC and DSC), nanoDSF, spectrophotometers and an Analytical Ultracentrifuge (UCA), and an optical biosensor based on surface plasmon resonance (SPR).

These techniques all aim to characterize molecular interactions (proteins, nucleic acids, sugars, lipids, small molecules, peptides, proteoliposomes, viral particles, etc...). They have in common the absence of the use of marked reagents, simplicity and speed of implementation. The complementarity of these techniques is widely referenced for the characterization of protein-ligand interactions

This platform is open to the academic and industrial research community.

The PINT platform is located on the 3rd floor of the green wing of the Faculty of Medicine

 

Microcalorimetry Pole

P.I. : François DEVRED

Equipment available:  3 microcalorimeters: ITC200, VP-ITC and VP-DSC allowing both isothermal titration calorimetry (ITC) and temperature scanning calorimetry (DSC) studies to be conducted, as well as the NT.Plex and Tycho nanoDSF.

The ITC makes it possible to characterize intermolecular interactions by obtaining their complete thermodynamic profile: affinity constant, stoichiometry, enthalpy and entropy.

The DSC and the nanoDSF make it possible to determine the thermostability (5-120 °C) of proteins, DNA, micelles... which gives indications on the tertiary structure of molecules and molecular interactions. The nanoDSF through the use of capillaries can also be used for quality control and to screen interactions.

In parallel with our service activity, we are conducting an R&D activity aimed at developing a new diagnostic method by calorimetric profiling of biofluids. In collaboration with INP Team 8 and the Timone Neuro-oncology Department (AP-HM) we are working on the development of a method for monitoring glioblastoma (Tsvetkov et al Oncotarget 2018). In partnership with the QARMA team of the Laboratoire Informatique et Systemes of Aix Marseille University and the company EURANOVA, both specialists in machine learning, we are developing a universal approach aimed at extending this method to other diseases of the nervous system (Alzheimer's, Parkinson's, multiple sclerosis, etc.).

 

Ultracentrifugation analytique Pole

P.I. : Pascale BARBIER

Equipment available : Beckman XLA Analytical Ultracentrifuge

Analytical UltraCentrifugation (UCA) allows the study of the behaviour of native macromolecules subjected to centrifugal force in solution. With the help of a detection system that measures the macromolecule concentration as a function of the centrifugation radius, we can perform two types of experiments that will lead to complementary informations:

The sedimentation velocity experiment mesures the sedimentation coefficient, the shape, the associative state (monomer, dimer, trimer,...) of a macromolecule. It can also highlight the formation of a complex and allows the determination of its affinity constant.

The sedimentation equilibrium mesures the molecular mass, the associative state (monomer, dimer, trimer,...) and the formation of a macromolecular complex.

 

Surface Plasmon Resonance pole

P.I. : Géraldine FERRACCI

Equipment available : Biacore T200, Biacore 3000 et ProteOn XPR36

SPR systems are designed to follow in real time interactions between free-labeled biomolecules. One reactant, the ligand, is immobilized on the sensor chip and the others interaction partners, named analytes, are injected at constant flow with a microfluidic circuit other the interface. The SPR principle is to follow mass changes on the surface of the sensor chip, induced by molecular complex formation and dissociation.

The use of real-time measurement allows to provide association (ka) and dissociation (kd) rate constant of the interaction and to deduce its affinity (KD). With an automated, parallelized and miniaturized mode of analysis, SPR optical biosensors also allow to define thermodynamic constants, the active concentration of an analyte, epitope mapping or study molecules in complex environments and perform micropurification and recovery of unknown partners.

 

Proteomic analysis CAPM pole

P.I. : Maya Belghazi, maya.belghazi@univ-amu.fr

Equipment available : Micro-HPLC Smart System BioRad ; Ultraflex II TOF-TOF Bruker ; Q-TOF Ultima Waters ; TECAN EVO 100 ; Nano UHPLC DIONEX 3000.

Mass spectrometry is a physical analytical technique that allows molecules to be detected and identified by measuring their mass and characterizing their chemical composition.

- Whole mass

- PMF or MSMS identification

- iTRAQ Quantification

- Sequencing de novo

 

=> Venez découvrir ces approches (et d'autres) en vous inscrivant à notre workshop MediM 2022

Partners: 

         

Non exhaustive list of PINT’s collaborators :

Marseille medical genetics, Inserm UMR U1251, Marseille (Marc Bartoli) - Institut de Chimie de Nice, UMR CNRS 7272, Nice (Rachid Benhida) - Centre européen de recherche en imagerie médicale, Marseille (Benjamin Guillet) - INP, Institut de NeuroPhysiopathologie, UMR CNRS 7051, Marseille (Francois Devred, Emeline Tabouret) - CRCM, Centre de recherche en cancérologie de Marseille, Inserm U1068, Marseille (Joseph Ciccolini, Eddy Pasquier) - Laboratoire d'Ingénierie des Systèmes Macromoléculaires,UMR CNRS 7255, Marseille (James Sturgis) - Grenoble Institut Neurosciences, Inserm UMR U1216, Grenoble (Isabelle Arnal) - Architecture et Fonction des Macromolécules Biologiques, CNRS UMR 7257, Marseille, (Sonia Longhi) -  Institut de Microbiologie de la Méditerranée, Laboratoire de chimie Bactérienne, CNRS UMR 7283, Marseille, (Romé Voulhoux) - Institut de Microbiologie de la Méditerranée, Bioénergétique et Ingénierie des Protéines, CNRS UMR 7281, Marseille, (Brigitte Gontéro)

Détails

Certification: 

PFNT is a facility labeled AMU.

 

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News

  1. INP attending first MOSBRI meeting in Paris

    On the 20-22 nd June, the first Molecular Scale Biophysics Research Infrastructure (MOSBRI) conference was held in Paris. Dahbia Yatoui from team 9 and Géraldine Ferracci from PINT went to this event, where Dahbia had the opportunity to present her work "Role of zinc-binding sites in Tau aggregation" on a poster.

  2. New publication : "Identification of the three zinc-binding sites on tau protein" from team 9 and PINT

    Team 9 and PINT are glad to see Romain La Rocca's main PhD paper is finally out in the International Journal of Biological Macromolecules.

  3. MEDIM 2022 - retour d'expérience

    Du lundi 24 au vendredi 29 avril s'est tenue, toujours avec succès, la deuxième édition de la formation MedIM (Méthodes d'étude des Interactions Moléculaires) co-organisée par la Plateforme INteractome Timone (PINT) de l’INP, la plateforme de Biologie Structurale de l’AFMB et les plateformes de l’IMM, toutes trois labellisées "Plateformes Technologiques du site d'Aix-Marseille".

  4. L'équipe GlioME et la plateforme PINT mobilisées lors de l'assemblée générale de l'association ARTC Sud

    Emmanuel Snacel-Fazy, doctorant dans l'équipe Gliome et  François Devred co-directeur de la plateforme PINT ont présenté leurs travaux lors de l'assemblée générale de l'Association pour la Recherche sur les Tumeurs Cérébrales (ARTC Sud).

  5. Recent works on Zinc binding to S100A1 and to NCS-1 published in Biomolecules and in  International Journal of Molecular Sciences respectively

    Team 9 (Cytoskeleton and Neurophysiopathology) and platform PINT (platform interactive Timone) are happy to see the almost simultaneous publications of two papers about about the impact of Zn2+ binding to NCS1 and to S100A1 respectively with their collaborators from Belozersky Institute of Physico Chemical Biology, Lomonosov Moscow State University, while the third one about Tau protein Zn2+ binding sites has been under review for already 3 months ... 

  6. Romain La Rocca giving a talk in EuroTau 2021 meeting

    Romain La Rocca, former member of INP team 9, shared the stage of EuroTau meeting in Lille's Grand Palais with Illana Gozes (Israel), Cláudio M. Gomes (Portugal), Amandine Grimm (Switzerland), Yann Fichou (France), Marina Rierola (Germany), Laura Vallés Saiz (Spain), Marie Galas (France) and Mahmoud Maina (UK / USA) during Session #3 chaired by Isabelle Landrieu & Yann Fichou.

    He presented his PhD work on the Three zinc binding sites of Tau and their role in tau aggregation and little bit of the perspectives Dahbia Yatoui is currently working on right now.

     

  7. INP attended Canceropole PACA 8th annual seminar

    Last week the 8th annual seminar of Canceropole PACA took place in the Palais des Congrés of Saint Raphaël. During these 2 days a great number of scientific presentations were given, including different aspects of INP research coming from team 8, team 9 and the platform PINT, during the poster sessions.

  8. INP first video abstract ?

    A couple weeks after the release of article "An AI-Powered Blood Test to Detect Cancer Using NanoDSF" authored by Tsvetkov Philipp*, Eyraud Rémi*, Ayache Stéphane, Bougaev Anton, Malesinski Soazig, Benazha Hamed, Gorokhova Svetlana, Buffat Christophe, Dehais Caroline, Sanson Marc, Bielle Franck, Figarella Branger Dominique, Chinot Olivier, Tabouret Emeline* and Devred François* in Cancers, the two first and two last authors of this interdisciplinary study participated in their first video abstract. Here is the result ... be kind 

     

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Job offers

Platform engineer : Conformational stability of proteins and protein ligands interactions

The Platform Interactome Timone of the Institute of Neurophysiopathology (INP) located at the Medical Faculty of AMU at Timone campus (Marseille) is seeking a dynamic and talented engineer with strong background in biophysical methods and protein biochemistry for participation in several interdisciplinary projects. Our unit uses biophysical techniques (Isothermal titration calorimetry, differential scanning calorimetry, differential scanning fluorimetry, dynamic light scattering, surface plasmon resonance etc.) to study the molecular mechanisms implicated in neurodegeneration and to develop new diagnostic methods.

The engineer will be involved in developing a novel universal diagnostic method based on denaturation profiles of human plasma samples that will be analyzed with the help of machine learning algorithms. Also, he/she will participate in the screening of a large library of inhibitors of protein-protein interactions to find new compounds that impact microtubular cytoskeleton.

The ideal candidate must have skills in biophysical methods, molecular biology, and biochemistry.
The position is opened for one year, renewable up to 2 years starting  from september 2021. Net wage : 1500-1800 net
Address : Plateforme PINT, Institut de NeuroPhsyiopathology, UMR 7051, Faculté des Sciences Médicales et Paramédicales, 27 Bd Jean Moulin, 13005 Marseille
website : https://inp.univ-amu.fr/en/facilities/pint

Applicants must email a CV, a statement of interest and contact details for 2-3 references to francois.devred@univ-amu.fr

Relevant publications:

2020     An AI-powered blood test to detect cancer using nanoDSF

2020     In silico molecular target prediction unveils mebendazole as a potent MAPK14 inhibitor

2019     Plasmatic Signature of Disease by Differential Scanning Calorimetry (DSC)

2018     Differential scanning calorimetry of plasma in glioblastoma: toward a new prognostic/monitoring tool