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About Membrane Protein Crystallisation, Grenoble, France

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Lipidic cubic phase (LCP) crystallisation is an alternative to standard crystallisation for membrane proteins. It allows membrane proteins to crystallise in a close to native and stable environment, in a lipidic bilayer. G-protein-coupled receptors (GCPR) protein crystallisation demonstrated the power of this approach; Cherezov V et al. High Resolution Crystal Structure of an Engineered Human β2-Adrenergic G protein-Coupled Receptor. As LCP is described as infinite 3D periodic structure, membrane proteins (MP) can freely diffuse laterally along the membrane. Diffusion is one of the important pre-crystallisation characteristic of the MP embedded into lipidic bilayer, which could be checked by Fluorescence recovery After Photobleaching (FRAP) technique available at HTMPC platform.

Instruments Available:

The platform is fully equipped to accomplish successful in meso crystallization (lipidic cubic phase, LCP) of membrane proteins. Robotic nanovolume dispensers (NT8 and Mosquito) screen 96 precipitant conditions with small amount of sample (5µl per plate) in few minutes. Our platform works with commercially available kits specialized in membrane protein crystallization. Automated imaging systems, RockImager 1500 and 1000 (Formulatrix), visualize crystallization drops with visible and UV lights. HTMPC is now part of the HTX Lab (EMBL) and benefits from CrystalDirect technology developed at the HTX Lab. CrystalDirect plate allows automated crystal harvesting and is well adapted to crystals grown in LCP drops.

LCP-FRAP measures membrane protein mobility in lipidic bilayer, which correlates with crystallisation in LCP.

User Guide

High Throughput MPC platform is fully equipped to accomplish successful crystallisation of your membrane protein. 

  • Robotic systems nanovolume dispensers (Mosquito and NT8) allow screening 96 precipitant conditions with small amount of sample (5-10 µl per plate) in less than 10 minutes. Our platform works with commercially available kits specialized in membrane protein crystallization 
  • Automated imaging systems, RockImagers (Formulatrix), visualize crystallization drops with visible and UV lights, which allow a better identification of protein crystals
  • The diffusion rate and mobile fraction of the membrane protein can be determined by FRAP (Formulatrix) that allows identifying the optimal conditions for LCP crystal growth

Instruct Centre

Instruct Centre FR2

IBS-ISBG

71 avenue des Martyrs

Grenoble

France

www.ibs.fr & www.isbg.fr

Membrane Protein Crystallisation, Grenoble, France

Contacts:

Florine Dupeux
Florine Dupeux
Institut de Biologie Structurale
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