NGFN-PLUS
Platform Proteomics
| Coordinator: | Prof. Dr. Katrin Marcus | |
| Institution: | Ruhr Universität Bochum | |
| Homepage: | funktionelle-proteomik.de |
Together with our cooperation partners of the integrated genome research network we are aiming to unravel the cause of Parkinson’s disease (PD). By understanding the etiology of PD it will be feasible to develop new drugs in the future to delay or even arrest disease progression.
Proteins exert diverse main functions such as building structural cellular components or performing molecular transport, facilitating chemical reactions and allowing signal transduction. By this, proteins meet criteria like “molecular machines” defining the function of a cell. We investigate the entirety and complexity of protein species within a cell or organism designated here as the proteome. In the Medizinische Proteom-Center of the Ruhr-University of Bochum (MPC) we apply diverse state-of-the art techniques which are able to display several thousands of proteins in parallel, termed as proteomics. The detection of regulated proteins is performed using a method called two-dimensional Fluorescence Difference Gel Electrophoresis (2D-DIGE). By this, we can separate thousands of protein species within one gel with concomitant measuring of their relative abundances. Afterwards, the protein identity can exactly be determined by mass spectrometry.
Together with our collaboration partners, we intent to compare the proteomes of disease patients samples versus those of healthy controls. In addition, we aim to investigate appropriate model systems, for example cell lines or mouse strains which bear mutations of hereditable forms of PD. Several data suggest alterations of the immune system to be causative for the development of PD. Therefore, in a further study we address this question by analysing blood samples with respect to auto-immune antibodies reacting to protein biochips.
All these approaches will yield differences in the protein composition between disease and healthy state pointing to mechanisms which lead to disturbed cellular functions. Additionally, disease-related altered proteins might serve as marker molecules for the diagnosis of Parkinson’s disease.
Two-dimensional Fluorescence Difference Gel Electrophoresis (2D-DIGE)
Description:
By applying an electrical field a rather complex mixture of proteins can be separated. In the first dimension the proteins arrange due to focusing on an acidic to basic membrane strip (IEF-PAGE). In the second dimension the pre-separated protein mixture is divided further according to the molecular weight of the proteins (SDS-PAGE). The result of the separation can be visualized by a silver staining (A). The detection of differences in protein abundance can be achieved by labelling the proteins of different samples with green, red or blue fluorescent dyes prior to mixing them together. After the gel run the proportions of the respective dyes can be measured and calculated (B).
Additional relevant Internet link:
Medizinisches Proteom Center
Further Coordinators:
Proteins exert diverse main functions such as building structural cellular components or performing molecular transport, facilitating chemical reactions and allowing signal transduction. By this, proteins meet criteria like “molecular machines” defining the function of a cell. We investigate the entirety and complexity of protein species within a cell or organism designated here as the proteome. In the Medizinische Proteom-Center of the Ruhr-University of Bochum (MPC) we apply diverse state-of-the art techniques which are able to display several thousands of proteins in parallel, termed as proteomics. The detection of regulated proteins is performed using a method called two-dimensional Fluorescence Difference Gel Electrophoresis (2D-DIGE). By this, we can separate thousands of protein species within one gel with concomitant measuring of their relative abundances. Afterwards, the protein identity can exactly be determined by mass spectrometry.
Together with our collaboration partners, we intent to compare the proteomes of disease patients samples versus those of healthy controls. In addition, we aim to investigate appropriate model systems, for example cell lines or mouse strains which bear mutations of hereditable forms of PD. Several data suggest alterations of the immune system to be causative for the development of PD. Therefore, in a further study we address this question by analysing blood samples with respect to auto-immune antibodies reacting to protein biochips.
All these approaches will yield differences in the protein composition between disease and healthy state pointing to mechanisms which lead to disturbed cellular functions. Additionally, disease-related altered proteins might serve as marker molecules for the diagnosis of Parkinson’s disease.
Two-dimensional Fluorescence Difference Gel Electrophoresis (2D-DIGE)
Description:
By applying an electrical field a rather complex mixture of proteins can be separated. In the first dimension the proteins arrange due to focusing on an acidic to basic membrane strip (IEF-PAGE). In the second dimension the pre-separated protein mixture is divided further according to the molecular weight of the proteins (SDS-PAGE). The result of the separation can be visualized by a silver staining (A). The detection of differences in protein abundance can be achieved by labelling the proteins of different samples with green, red or blue fluorescent dyes prior to mixing them together. After the gel run the proportions of the respective dyes can be measured and calculated (B).
Additional relevant Internet link:
Medizinisches Proteom Center
KTT
MEDIA
CURRENT
JOBS
NGFN- MEETING
NGFN1&2
LINKS