RIBOSOME-SECYEG · Crystallographic studies of the complex between the prokaryotic ribosome and the SecYEG translocon
FP6 — Marie Curie Actions (Human Resources and Mobility)
- Duration
- 2007-11-07 → 2009-11-06
- EU contribution
- €177,233
- Participants
- 1
- Scheme
- EIF
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Results in brief
Final Activity Report Summary - ribosome-SecYEG (Crystallographic studies of the complex between the prokaryotic ribosome and the SecYEG translocon)
A reliable protocol for obtaining satisfactory amounts of translation-arrested e.coli ribosomes that were suitable for crystallisation and co-crystallisation was established. In order to achieve this goal, several problems regarding quantity, stability and uniformity of the ribosomes had to be overcome. Extensive crystallisation screening produced numerous crystallisation conditions under which single macrocrystals of translation-arrested ribosomes could be obtained reproducibly. The initial X-ray diffraction tests of these crystals showed promising results and highlighted the potential for further improvement.
Data: CORDIS, © European Union
Project objective
We propose to crystallize the complex of the prokaryotic ribosome or its 50S subunit with the SecYEG translocon. Two SecYEG heterotrimers bind to the ribosome at the end of the nascent polypeptide tunnel to form a transmembrane channel which is part of a translocase complex that allows for secreted and membrane embedded molecules to be transcribed through or into the membrane bilayer respectively.Building on previously established expression and complex formation protocols that allowed the analysis of this complex via electron microscopy we will proceed to crystallize the complex. We will rely on a multi-pronged approach involving ribosomes and SecYEG proteins from various organisms, which are able to form heterologous complexes, as well as engineered/truncated versions of the SecYEG proteins.Our goal is to solve the crystal structure of this complex at atomic resolution to shed light on the exact mechanisms of one of the most basic and important biological processes. Questions we hope the structure will help us address are how the channel is opened and closed, how membrane proteins are embedded into the bilayer from the channel and how the membrane seal is maintained throughout the whole process.The proposed project contributes to the development of x-ray crystallography of macromolecular assemblies and membrane proteins which are both regarded as future focal points of structural biology. As such it is valuable for European research and also provides excellent future perspectives for the researcher.The techniques involved in the project complement the skill-set of the researcher well and will leave him with well-rounded crystallographic expertise focused on two of the most promising areas in his field of research. Furthermore, funding this project would al low the researcher, who is from an objective 1 less-favoured region, to return to Europe from his current position in the United States.
Original text from CORDIS.
Participants
- EIDGENOSSISCHE TECHNISCHE HOCHSCHULE ZURICH · ZURICHCoordinatorCity levelSwitzerland
Links
Data: CORDIS, © European Union