PINT

Scientific supervisor: 
Description: 

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

 

=> notre workshop

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.

 

News

  1. dianette akta Geraldine biacore
    PINT new acquisitions: Biacore T200 & AKTA pure

    The Platform INteractome Timone - PINT is pleased to announce the arrival of a brand new Surface Plasmon Resonance Biacore T200 (Cytiva, ex GE Healthcare) designed to follow in real time interactions between free-labeled biomolecules from ions to viruses. The use of real-time measurement allows to provide association (kon) and dissociation (koff) rate constant of the interaction and to deduce its affinity (KD).

  2. INP teams awarded MIC Cancer Inserm - ITMO Aviesan 2020 grant
  3. MedIM 2020 : First inter-platform Interactome workshop

    INP platform PINT together with AFMB and IMM are glad to announce their first inter-platform interactome workshop entitled "Methods to study Molecular Interactions (Biomedical applications)" that will take place from monday december 7th to friday 11th 2020. 

    Looking to confirm the interaction between a drug and its target?
    Wanting to characterize the interaction between a receptor and its ligand?

  4. Nanotemper Technologies 1st Virtual Symposium 

    Romain La Rocca, third year PhD student in INP team 9 (Cytoskeleton and Neurophysiopathology), will present a part of his thesis work during the 1st NanoTemper Virtual Symposium on June 30th from 9 to 12h30. NanoTemper annual seminars bring together the researchers using NanoTemper instruments, or eager to discover new technologies and in different fields of research. During this event, Romain will show how to use the nanoDSF technology to study the quaternary structure of tubulin. 

  5. PINT new acquisition: Tycho™ NT.6 for instant protein quality control

    The Platform INteractome Timone - PINT is pleased to announce the arrival of the Tycho™ NT.6, the new nanoDSF instrument by Nanotemper. The Tycho is designed to characterize the quality of any protein within minutes, using only 10 µL of sample. This fast analysis is critical for choosing the batches, buffers, or conditions by comparing the relative stability of each sample along every step of an experiment, thus allowing an efficient assay and purification workflow development.

    Since it requires just 10 µL of sample at a concentration anywhere from 0.01 to >200 mg/mL, it's not even necessary to dilute samples. 

  6. poster prize at ARBRE MOBIEU 4th plenary meeting
    Romain La Rocca won best poster prize at ARBRE-MOBIEU 4th plenary meeting in Prague

    Romain La Rocca, third year PhD student in INP team 9 (Cytoskeleton and Neurophysiopathology), recently won the the best poster prize at the 4th ARBRE-MOBIEU pleanry meeting in Prague on February 24, 25 and 26.

    ARBRE MOBIEU pleanry meeting is a European Biophysics Congress that brings together many researchers in Europe from various fields, all of whom have in common the use of biophysical tools. On this occasion, Romain presented his thesis work on the effects of zinc on the Tau protein using NMR, ITC, DLS among other biophysical techniques.

  7. Philipp Tsvetkov receiving its GEFLUC grant in Hotel de la Région - 04th February 2020
    Philipp Tsvetkov received a 29 000 € grant from the GEFLUC
    • The Emergence project entitled “New high throughput screening method for anticancer agents” carried out by Philipp Tsvetkov (first from the right) was selected by the Canceropole PACA to be funded by the GEFLUC (groupement des entreprises françaises dans la lutte contre le cancer).
  8. "Alzheimer's disease: a year of hope" (La Marseillaise - January 2020)

Pages

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 3 years starting as early as January 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