-->
Service

Measuring protein structural dynamics

Single molecule FRET allows the measurement of distances and distance changes between two fluorescent molecules in a biomolecular complexes, down to the Angstrom and the microsecond timescales. smFRET measurement can be performed in vitro:
  • On diffusing molecules, in a confocal geometry, to measure distances and fast conformational changes
  • On immobilized molecules to measure association / dissociation kinetics and conformational changes in the seconds timescales

All projects are run as collaborations. Please This email address is being protected from spambots. You need JavaScript enabled to view it. for help choosing the most appropriate approach. We also provide advice to design your experiment and support with fluorescent labeling and data analysis. After a meeting, we can run a few experiments to assess the feasibility of the approach. Then a collaboration scheme is defined between the team and the facility. Our R&D division can also implement specific adaptations for your project if needed.

 

FRET

Measurements of distances changes within E. Coli and MTB transcription complexes using single molecule FRET (from Vishwakarma et al.)

 

Selected publications:

Quast R.B., Fatemi F., Kranendonk M., Margeat E.*, Truan G.* Accurate Determination of CPR Conformational Equilibrium by smFRET using Dual Orthogonal Non-Canonical Amino Acid Labeling, ChemBioChem, 2019, 20(5):659-666

Vishwakarma RK, Cao AM, Morichaud Z, Perumal AS, Margeat E, Brodolin K, Single-molecule analysis reveals the mechanism of transcription activation in M. Tuberculosis. Sci Adv, 2018

Ait-Bara S, Clerte C, Declerck N, Margeat E. Competitive folding of RNA structures at a termination / antitermination site. RNA, 2017

Ait-Bara S, Clerte C, Margeat E. Single-Molecule FRET Characterization of RNA Remodeling Induced by an Antitermination Protein, Methods Mol Biol, 2015

Olofsson L, Felekyan S, Doumazane E, Scholler P, Fabre L, Zwier JM, Rondard P, Seidel CA, Pin JP, Margeat E. Fine tuning of sub-millisecond conformational dynamics controls metabotropic glutamate receptors agonist efficacy, Nat Commun, 2014

 

Equipement

SmFRET TCSPC

 

TIRF / 3D MFM

       
smFRET
Read more ...
 
Read more ...
       

Connexion