Biochemistry - Protein analysis
The Central Core Facility of Protein Analysis comprises the proteome activities of the Institute of Infection Biology. The classical proteome analysis combining two-dimensional electrophoresis (2-DE) with mass spectrometry resulted in a proteomic database containing several thousand identified proteins of microorganisms, eukaryotic cells and tissues. Peptide mass fingerprinting with MALDI-MS or ESI-MS and protein sequencing in cooperation with the Technical University (Edman degradation) and the Max-Delbrück Center (MS/MS) revealed posttranslational modifications and by de novo sequencing not predicted genes in the mycobacterial genome. Complementary proteome technologies are used and developed like immunoproteomics, isotope coded affinity tag mass spectrometry, post-translational modification proteomics and subproteomics.
The structure of the Central Core Facility allows cooperating at different levels of intensity:
- Service level
- Use of technology
- Planning and performance of proteomic investigations
- Sample preparation
- Gels up to the size of 30 x 40 cm
- Several detection methods
- Blue native electrophoresis
- In-gel or in–solution digestion with several proteases
- Identification of proteins using MALDI MS +/- MALDI MSMS or nano-LC MALDI MSMS
- Quantification using peptide-ITRAQ-Labelling
2-DE database (in cooperation with the Bioinformatics Core Facility): tools for the construction of a 2-DE database (http://www.mpiib-berlin.mpg.de/2D-PAGE/). Access to N-terminal sequencing (Edman degradation), ESI-Ion Trap MS, data dependent MS.