Institute of Technology, Chemistry and Biology (ITQB- UNL), Oeiras, Portugal
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Membrane proteins
(MP) are poised at the interface between the cell and the surrounding environment,
and perform key biological functions such as energy transduction, ion transport
and regulation, molecular recognition and response. Thus, they are major targets
for many pharmacological compounds. Our understanding of the function of this
broad range of proteins is highly circumscribed by a lack of structural information,
due to limitations normally associated with the production of large amounts
of membrane proteins, either native or recombinant, and the difficulty in obtaining
crystals suitable for X-ray diffraction.
Membrane proteins,
containing both hydrophilic and hydrophobic domains, are more difficult to isolate
than water-soluble proteins. The native membrane surrounding the protein must
be disrupted and replaced with detergent molecules without causing any denaturation
of the protein itself.
Despite the importance of MP in cell function, a cursory examination of the Protein Data Bank (PDB) will reveal that very little information is available on membrane protein structure. The number of soluble protein structures in the database exceeds the number of membrane protein structures by approximately 1000:1! This represents a huge deficit of information about MP structure. The reason behind this extraordinary deficit of membrane protein structures is largely due to insolubility of membrane proteins in aqueous solutions, due to their amphipathic nature. Detergents, which mimic the lipid bilayer, are used in order to solubilize MP in aqueous environment. However, detergents tend to inhibit crystallization, and crystals are required for X-ray Crystallography structure determination. Therefore, the limitation normally associated with the production of large amounts of membrane protein, either native or overexpressed and the difficulty in obtaining well ordered crystals for X-ray diffraction are the major obstacles to 3D structure determination of proteins associated with the membrane.
In recent years, structural membrane protein research is rapidly evolving with an increasing number of structures being determined. Since new approaches are being developed within this field, we think it is extremely important to organize a course that will provide young scientists the opportunity to learn of the latest developments in a range of techniques relevant to membrane protein production and crystallization from international experts in the field.
The course will focus on the expression, purification and crystallization of membrane proteins. A wide range of methodologies will be covered, namely, improvements of overproduction of membrane proteins both in their native environment or refolded from inclusion bodies, procedures for overexpression and purification, availability of new detergents, and antibody fragment mediated crystallization of membrane proteins.
The 6 day course
will include lectures in the morning and tutorial/practical sessions in the
afternoon. Participants will have the opportunity of presenting a poster, some
of which will be selected for oral communication. Extended discussion among
the speakers and participants will be encouraged through poster discussion sessions,
during lectures and tutorials and in a final round-table discussion with students
and speakers. During the "hands-on" practical sessions, the students
will be able to overexpress and purify a selected membrane protein, using several
chromatographic steps (FPLC equipment is available). The crystallization screenings
will be held on a state of the art nanoliter crystallization robot, an essential
tool for a completely automated crystallization approach and crucial for the
number of conditions to be tried and the amounts of membrane proteins usually
available. Training the widest possible number of students in this technology
is fundamental for the advance and development of membrane proteins crystallization
and structure determination.
Venue:
Institute of Technology, Chemistry and Biology (ITQB- UNL), Oeiras, Portugal
19 - 24 September, 2005
Confirmed Speakers:
Prof. So Iwata,
UK
Membranes. A new era in membrane protein studies
Crystallization and Crystallography of membrane proteins
Dr. Bernadetted
Byrne, UK
Membrane protein expression
Membrane protein purification and crystallization
Dr. Edmund Kunji,
UK
Lactococcus lactis as host for overproduction of functional membrane proteins
Linking electron and X-ray crystallography of membrane proteins
Prof. Thue Scwartz,
Denmark
Structure and function of 7TM receptors
Prof. Gunnar von
Heijne, Sweden
Membrane protein biosynthesis and assembly
Prof. Peter Henderson,
UK
Principles of membrane transport
How to catch and characterize membrane transport proteins
Dr. Carola Hunte,
Germany
Antibody fragment mediated crystallization of membrane proteins
Membrane protein purification for structural studies
Importance of phospholipids for structure and function of membrane proteins
Dr. Kaspar Locher,
Switzerland
Homolog screening in membrane protein crystallization: the ABC transporter case.
Registration is now closed.