STRUCTURE OF YIDC · Structural characterization of YidC, a novel component of the membrane proteins translocation machinery
6РП — Действия „Мария Кюри“
- Период
- 2006-11-01 → 2008-10-31
- Финансиране от ЕС
- 148 020 €
- Участници
- 1
- Схема
- EIF
Линиите свързват координатора с партньорите.
Накратко на български
Протеинът YidC и неговата структура се изследват чрез анализи на бактериита E. coli. Разбирането на този механизъм помага да се разбере как се изграждат и подреждат протеините в мембраните на всички живи клетки.
Кратко обяснение, генерирано от езиков модел по текста на CORDIS. Оригиналът е по-долу.
Резултати накратко
Final Activity Report Summary - STRUCTURE OF YIDC (Structural characterisation of YidC, a novel component of the membrane proteins translocation machinery)
Biogenesis of inner membrane proteins is of central importance for all cells. In bacteria it requires targeting and insertion factors such as the signal recognition particle and the Sec translocon. YidC is an essential membrane protein, recently described as a component of the insertion machinery. Members of the YidC family (YidC/Oxa1/Alb3) exist in all three domains of life where they have multiple functions in the integration and assembly of a large variety of membrane protein complexes. In this research project, we aimed to gain insight into the structure and function of YidC using a multidisciplinary approach. As a first essential step of the study, a rational optimisation of the cloning, production and purification protocols of E. coli YidC was established, and later successfully applied to homologous proteins. The oligomeric state and stability behaviour of the proteins were characterised in various detergent solutions. The interaction networks between YidC and the SRP and Sec translocon components were explored using complementary biophysical techniques. Broad range crystallisation screening was set up and yielded to microcrystals. The crystallisation of membrane protein represents a challenging work but many strategies were developed and planned to enhance the changes to success. The first high resolution structural data on the YidC protein family were obtained on the large periplasmic domain of E. coli YidC. The structure was determined at 1.8A resolution and revealed an unexpected fold and ligand binding site. This first breakthrough in the structure determination of the full length protein set the basis for structure function relationship studies.
Текст от CORDIS, на английски · Данни: CORDIS, © Европейски съюз
Цел на проекта
Understanding membrane protein biogenesis is of central importance for modern society as membrane proteins are not only crucial for transport or acting as receptors, they also comprise a major drug target for phamaceutical industry. In E. coli membrane protein biogenesis requires components of the well- described SRP and Sec pathways for targeting to and insertion into the membrane. However, a clear picture of the integration of membrane proteins into the lipid bilayer has not yet emerged. Recently, a membrane protein (YidC) essential for viability of E. coli has been identified. Members of the YidC family exist in all three domains of life where they have multiple functions in the integration and assembly of a large variety of membrane protein complexes.The host institution has established an efficient expression and purification protocol for YidC which allows now for the detailed structural and biochemical characterization of YidC alone as well as in complex with functionally relevant partners. Several detergents will be systematically screened for their influence on the oligomeric state of YidC as well as on its function. The interactions of YidC with the SRP system will be analysed and potential binding partners e.g. the SRP receptor, will be included in t he biochemical and structural studies. Besides E. coli, thermophilic bacteria will be used as source for YidC and also chimeras will be tested for structure analysis.The results obtained will not only contribute to the molecular details of membrane protein biogenesis, but will also add to a rational optimisation of membrane protein production by engineering the membrane insertion pathway. Due to the combination of biochemical and biophysical methods this project represents an excellent opportunity to strengthen my background in membrane protein crystallography and to complement my previous expertise with methods from molecular biology, biochemistry of protein complexes and electron microscopy.
Оригинален текст от CORDIS (на английски).
Участници
- RUPRECHT-KARLS-UNIVERSITÄT HEIDELBERG · HEIDELBERGКоординаторГермания
Връзки
Данни: CORDIS, © Европейски съюз