Membrane Proteins - the rocky road from gene to crystal
19-24 September 2005
Oeiras, Portugal

Organisers:

Margarida Archer Frazao, ITQB-UNL, Oeiras, Portugal
So Iwata,Imperial College, London, UK

Draft Report

Summary

Although ~30% of proteins encoded by genomes are predicted to be Membrane Proteins (MP), not many three-dimensional MP structures are available. Our understanding of the function of this broad range of proteins is highly circumscribed by a lack of structural information, due to the difficulty in obtaining crystals suitable for X-ray diffraction and also the limitation normally associated with the production of large amounts of membrane proteins. The interest on MP is emerging very quickly over the last years and the number of MP structures solved is increasing. Since many efforts are being developed within this field, the workshop was extremely important for covering the recent results in the area of expression, purification, crystallization and expression of membrane proteins.

The scientific programme focused on the methodology involved in the expression, purification and crystallisation of membrane proteins. Several state-of-the art techniques were presented by experts in these areas.

Scientific Content

The course lasted for 6 days and included lectures in the mornings and tutorial, practical and poster sessions in the afternoon. The lectures covered a wide range of topics related with membrane proteins, namely overexpression, purification, crystallization, and tools for structural and functional analysis of membrane proteins. There was a good balance between lectures covering basic topics and state-of-the-art methodologies, and lectures focusing on more specific subjects with presentation of some case studies. Since most of the participants had some experience on the area, they raised many questions during the lectures, so they were all very participative.
The lectures took place in the auditorium and were open to researchers with interest on the field, so all the 11 Portuguese students that applied to participate in the course were able to attend the theoretical lectures. We also had some students and more senior scientists from ITQB coming to the lectures.

The students were divided in 2 groups in the afternoon - one group for the laboratorial practical session, and the other group for the tutorial and poster session. Extended discussion among the speakers and participants took place during tutorial and poster session discussions. There were 3 laboratorial sessions - handling of membrane proteins, screenings and crystal optimization, and crystallization with lipidic cubic phases (demonstration). See the scientific programme for more details.

The course evaluation done by the students after the course was very positive - they learned a lot, had plenty of time for informal discussions, which was good to raise some questions and ask about specific problems they were facing during their research projects. Some suggested a bit shorter course (4 to 5 days, instead of 6). Hotel and social events were most appreciated. Participants had time to learn science and have fun!

Assessment of the results and impact of the course

The scientific level of the course "Membrane Proteins: the rocky road from gene to structure" was of high quality. The broad content of the lectures covered some more basic principles inherent to membrane proteins but also more advance techniques. Several speakers presented novel developments being done on the field and showed unpublished results during their lectures. Since all the topics were related with different approaches to the membrane protein field, the course allowed an updated discussion of what is going on and the efforts being developed on where to go next.

There was also time for practical sessions related with different techniques used for screenings, growth and optimisation of membrane protein crystals. The poster session and tutorials were very useful for the participants, since they had the opportunity to interact with other students and speakers, to exchange ideas and ask for suggestions on how to proceed in their future work. This was also fruitful because most of the students had some previous experience with membrane proteins, either on the level of expression, crystallization or characterization.

The course enabled the get together of speakers and students coming from different laboratories and institutes, mostly from all over Europe. To be able to contact personally with other people in the field is very important and potenciate future collaborations and joint projects. In fact, some participants are members of the European Membrane Protein Consortium E-MEP, and with this workshop many information was exchanged between them and non- EMEP members. As a result of the course, the participants created a mailing-list, so people could exchange ideas, ask questions about specific problems they have concerning their research work and positions available related to the field.

Programme
Monday 19 September
09:00-10:00 Material distribution
10:00-11:00 Crystals at all Peter Henderson, UK
11:00-11:20 Coffee break
11:20-12:20 Membrane protein expression Bernadette Byrne, UK
12:20-13:20 Large scale Production of G-protein coupled receptors in Pichia pastoris Christoph Reinhart, Germany
13:30-15:00 Lunch
15:00-18:00 Practical 1: Handling membrane proteins - Group A
15:00-16:20 Poster session - Group B
16:30-18:00 Tutorials - Group B Kaspar Locher, Switzerland, Edmund Kunji, UK, Christoph Reinhart, Germany
18:00 Welcome Reception (sponsored by Oeiras City Hall), Palácio Marquês Gardens

Tuesday 20 September
09:00-10:00 Membrane protein purification and crystallization Bernadette Byrne, UK
10:00-11:00 Membrane protein biosynthesis and assembly - part I Gunnar von Heijne, Sweden
11:00-11:20 Coffee break
11:20-12:20 Membrane protein biosynthesis and assembly - part II Gunnar von Heijne, Sweden
12:20-13:20 Principles of membrane transport Peter Henderson, UK
13:30-15:00 Lunch
15:00-18:00 Practical 1: Handling membrane proteins - Group B
15:00-16:20 Poster session - Group A
16:30-18:00 Tutorials - Group A Eva-Peyroula, France, Christoph Reinhart, Germany, Gunnar von Heijne, Sweden
20.00 Welcome dinner, Kais da Adega

Wednesday 21 September
09:00-10:00 Lactococcus lactis as host for overproduction of functional membrane proteins Edmund Kunji, UK
10:00-11:00 Homolog screening in membrane protein crystallization Kaspar Locher, Switzerland
11:00-11:20 Coffee break
11:20-12:20 Biochemical and structural work on the ABC transporters Kaspar Locher, Switzerland
12:20-13:20 Crystallization of the ADP/ATP carrier Eva-Peyroula, France
13:30-15:00 Lunch on Poster session (ALL)
15:00-18:00 Free Afternoon

Thursday 22 September
09:00-10:00 How to catch and characterize membrane transport proteins Peter Henderson, UK
10:00-11:00 Novel developments in lipidic cubic phase crystallization Eva-Peyroula, France
11:00-11:20 Coffee break
11:20-12:20 Antibody fragment mediated crystallization of membrane proteins Carola Hunte, Germany
12:20-13:20 Expression of membrane proteins in eukaryotic systems Christoph Reinhart, Germany
13:30-15:00 Lunch
15:00-18:00 Practical 2 - Crystallization of membrane proteins - group A
15:00-16:30 Demonstration - Lipidic cubic phase crystallization - group B
16:30-18:00 Tutorials - group B Peter Henderson, UK, Edmund Kunji, UK, Thue Scwartz, Denmark
20.00 Congress Dinner - Sponsored by Oeiras City Hall

Friday 23 September
09:00-10:00 The structure and Function of ClC chloride channels Raimund Dutzler, Switzerland
10:00-11:00 Importance of phospholipids for structure and function of membrane proteins Carola Hunte, Germany (CH)
11:00-11:20 Coffee break
11:20-12:20 Structure and function of 7TM receptors -I Thue Scwartz, Denmark
12:20-13:20 Crystallization and Crystallography of membrane proteins So Iwata, UK
13:30-15:00 Lunch
15:00-18:00 Practical B2Laboratorial session on Crystallization screenings on nanoliter robot and crystal optimisation
15:00-16:30 Poster session A
16:30-18:00 Tutorials A Raimund Dutzler, Switzerland, Carola Hunte, Germany, Christoph Reinhart, Germany

Saturday 24 September
09:00-10:00 Structure and function of 7TM receptors -II Thue Scwartz, Denmark
10:00-11:00 Tools and experiences in membrane protein crystallography Raimund Dutzler, Switzerland
11:00-11:20 Coffee break
11:20-12:20 Linking electron and X-ray crystallography of membrane proteins Edmund Kunji, UK
12:20-13:20 NADH dehydrogenase: the bumpy road from the crystal to the structure Margarida Archer, Portugal
13:30-15:00 Lunch
15:00-18:00 Round Table Students/ Speakers