| Description |
MePNet G-protein coupled receptor project coordinated by Bio-Xtal (www.bioxtal.com). Since then this project has taken on larger dimensions and will soon be launched in September.
The Objectives:
The MePNet Project will select the best validated, most advanced methods to express 100 GPCRs in 3 expression systems (E-coli, Pichia Pastoris, Similiki Forest Virus infected cells). On the basis of the production level, the best targets will be selected and only a limited number of receptors will be chosen for large-scale production, refolding, solubilization, purification & crystallization. The objective of this program is to attempt to develop, evaluate and improve all the tools used in the expression, stabilization and purification of GPCR targets.
Our aim is to obtain at least 5mg of pure functional protein suitable for crystallization trials on several studied targets. For those for which a greater amount can be obtained, as well as crystals, our ultimate goal is to solve their structure at the end of the program.
Our decision to start this project with 100 targets responds to three objectives:
(1) to establish a real statistic evaluation of the tested tools and protocols;
(2) to have the possibility to keep only the best evolving targets through the different stages;
(3) to conceive a high throughput approach suitable for many stages of GPCR studies.
The Academic teams involved:
· Dr. Franc PATTUS & team (Strasbourg, France): Expression systems, membrane protein biochemistry and biophysics;
· Dr. Hartmut MICHEL & team (Frankfurt, Germany): Molecular biology of GPCRs, membrane protein crystallography;
· Dr. Imgard SINNING & team (EMBL, Heidelberg, Germany): Membrane protein crystallography;
· Dr. Erik HUIZINGA & team (Utrecht, The Netherlands): Refolding from inclusion bodies, crystallo-genesis.
The expected results and milestones:
· Collection of cDNAs, clones, membrane fragments, functional assay protocols, ligands, and GPCR studies related protocols and materials, for both successful and unsuccessful experiments performed within and outside the network.
· Evaluation of effectiveness on the three main expression systems.
· Setting up of:
(1) stabilisation compounds for solubilisation, refolding, and crystallisation purposes
(2) quality control tools,
(3) high throughput technologies for cloning, quality tests, refolding, purification and crystallisation.
· 3D structures for several receptors
· Database released to the industrial partners.
The originality of this project lies in its attempt to consolidate as much information and material as possible from supporting pharmaceutical companies and academic laboratories, and create a database available for the network and supporting laboratories throughout the project.
The project already counts among its participants more than forty leading pharmaceutical companies and academic teams worldwide, including:
3D Pharma, American Home Products, Astra Zeneca, Novartis, Novo Nordisk, Roche, Sanwa Kagaku, Takeda….
If you are interested in participating in this project, even on a one-year trial basis, we invite you to visit the MePNet website (www.mepnet.org), where, under the heading « New Industrial Participants » you will find the details concerning the conditions for participation, how to benefit from an evaluation entry, what you will have access to, and how much this costs.
If you decide to participate, you are also cordially invited to attend the bi-annual general meeting of MePNet to be held in Strasbourg (France) on September 27th, 2001 at the:
CNRS, Ecole Supérieure de Biotechnologie de Strasbourg
Rue Sébastien Brandt,
67400 Illkirch
France.
At this meeting you will have the possibility to meet the current participants and the scientists involved, as well as find out about the techniques to be used and make your own contribution.
The agenda of activities for the day is as follows:
Thursday, September 27th 2001 (Morning):
9:00 - 10:20 - Presentation of the project organization, publishing rules, intellectual property and financial issues.
10:20 - 10:40 - Coffee break
10:40 - 12:00 - Scientific planning: means, milestones, database issues.
12:00 – 13:30 – Lunch Break
Thursday, September 27th 2001 (Afternoon):
13:30 - 14:30 - E-coli expression & quality control, refolding, and result analyses
14:30 - 15:45 - Pichia P. and Semliki FV expression & quality control, result analyses
15:45 - 16:00 - Tea break
16:00 - 17:30 - High throughput dimension of the expression, solubilization, refolding and functional assays protocols
If you need any further information or have any questions, please do not hesitate to contact us. We look forward to hearing from you soon.
Contact: info@mepnet.org
Web site: http://www.mepnet.org
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