Verwendbarkeit des Modules
- Master Pharma-Biotechnologie (Pflichtmodul) im 1. und 2. Semester
- Potential of biotechnological application of design proteins
- Proteins as pharmaceutical product-advantages and problems of therapeutically used proteins
- Function and construction of proteins
- Structure of proteins, protein-topology, data banks of primary structure, biocomputing of secondary and tertiary structures
- Homology-modelling, 3D-Imaging with viewer programmes based on sequence data
- Prediction of secondary structure elements and membrane-targeting domains
- Evaluation of precision of biocomputing-structural programs
- Properties of amino acids as principal units of protein engineering
- Molecular interaction of secondary structure elements of proteins
- Engineering of membrane proteins and receptors
- Molecular interaction of proteins with DNA: HLH- and Zn-finger domains
- Stability and denaturation, the problem of Prions
- Engineering of functional protein folding domains (Rhotekin-GST-Binding protein as molecular tool)
- Homology-modelling (superantigenes as example)
- Engineering of protein vaccines (pertussis toxin)
- Engineering of protein toxins as novel molecular tools (CNF1 and CNF-Y for cell signaling studies)
- Planning and protocol for amino acid exchange, insertion and deletion at any locus in proteins incl. special application of PCR-technology, primer-less methods
- Post-translational modification as engineering problem
- Protein targeting (antibodies etc.) and the current development of novel molecular therapies
Practical course Protein-Engineering:
All experiments were developed and performed under involvement of students in cooperation with the Institute for Molecular Cell Biology (Director Prof. Dr. R. Wetzker) and partially with the pharmaceutical company Serono (Geneve, Switzerland) and the Oncological Research Laboratory of the Friedrich-Schiller-University (head Dr. J. Clement).
Using a novel therapeutic targeting protein, PI3Kinase, the full potential of protein engineering will be demonstrated for pharmaceutical application. To illustrate this recent development all students will get a copy of the paper: R. Wetzker und C. Rommel. Phosphoinositide 3-kinase as targets for therapeutic intervention. Current Pharmaceutical Design 10, 1915-1922 (2004).
For further theoretical and practical support of the practical master courses contracts are planned between the Forschungszentrum für Molekulare Biomedizin and the Onkologisches Forschungslabor am Klinikum der FSU. They will enable the constant offer of the highest possible level in future education in this field of biotechnology.
- 3D-imaging using PI3-Kinase as example: Labelling of the active center and of the protein-interaction domain with the Ras-GTPase
- Sequence based modelling of secondary structures with computer programmes (PCGENE, DNAsis), comparison of the prediction with X-ray structural data
- Homology analysis with free internet programmes (preferably via ncbi des NIH), comparison of alignments of superantigens with different programmes
- Evaluation of the biocomputing structural programmes (Ras-GTPase, superantigen SPEC)
- Expression constructs with the signalling molecule PI3-Kinase (human) for higher cells
- Site-directed mutagenesis in the active center of the kinase (i. e. application of K/R-mutants)
- Insertion of the membrane-anchoring motif CAAX as an example to manipulate membrane targeting
- Expression-constructs functioning in human cells
- Transfer of constructs in human target cells, protein engineering of cellular monitoring systems, fluorescence monitoring with expression systems for YFP- und GFP-fusion proteins
- Overexpression of the PI3-Kinase WT/Engineering-kinase in a kinase-free cellular background
- Test of the endocytotic cellular function under expression of the engineering-constructs
- Test of endozytosis of ferromagnetic nanopartikel under expression of protein-engineering constructs
Students learn about the potential of protein design and construction in biotechnology and pharmaceutical application. The main focus will be novel development of protein products and diagnostics. Profound knowledge about protein structure will be communicated as prerequisite for all protein engineering work. The unlimited potential of protein engineering will be demonstrated theoretically and also as planning and established work protocols: Targeting via exchange, deletion, insertion and fusion of one or any number of amino acid residues resp. peptides for all therapeutic and diagnostic applications. Special methods like PCR without primers etc. will be communicated.
The practical course is the continuation and necessary completion of the laboratory course Genetic Engineering and Molecular Cell Biology. It is designed as integrated practical course with that of Molecular Cell Biology. It will respond to the rapidly growing demand for graduates especially trained in this important field of biotechnology. The Students experience the direct correlation between the current developments of protein engineering with functional analysis in higher cells: Identification of novel targets, drug design, development of molecular tools for diagnostics and therapy. Biocomputing will be performed concerning 3D-imaging of protein molecules, protein-protein-interaction, labelling of selected targeting-amino acid residues, sequence-based analysis of secondary and tertiary structures, homology-modelling and alignment-analyses.
Lehr- und Lernformen
|Teil 1||Teil 2|
Kenntnisse der Molekularen Zellbiologie.
Voraussetzungen für die Vergabe von ECTS Credits
Schriftliche Prüfungsleistung (90 min.), Laborschein
Skript und Praktikumsanleitung im Intranet als pdf-Datei
- Lehninger: Biochemie, Springer, 2001.
- Parekh, Rohlff: Post-translational modification of proteins and the discovery of new medicine, Current Opinion in Biotechnology, 1999.
- Gibbs, Ras: C-terminal processing enzymes – New drug targets, Cell 65, 1999.
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