551-0438-00L  Protein Folding, Assembly and Degradation

SemesterAutumn Semester 2015
LecturersR. Glockshuber, E. Weber-Ban
Periodicitysemester course
Language of instructionEnglish
CommentNumber of participants limited to 7.


AbstractStudents will carry out defined research projects related to the current research topics of the groups of Prof. Glockshuber and Prof. Weber-Ban. The topics include mechanistic studies on the assembly of adhesive pili from pathogenic bacteria, disulfide bond formation in the bacterial periplasm, ATP-dependent chaperone-protease complexes and formation of amyloid deposits in Alzheimer's disese.
ObjectiveThe course should enable the students to understand and apply biophysical methods, in particular kinetic and spectroscopic methods, to unravel the mechanism of complex reactions of biological macromolecules and assemblies in a quantitative manner.
ContentThe students will be tutored in their experimental work by doctoral or postdoctoral students from the Glockshuber or Weber-Ban group. In addition, the course includes specific lectures that provide the theoretical background for the experimental work, as well as excercises on the numeric evaluation of biophysical data, and literature work.

Participation in one of the following projects will be possible:

Projects of the Glockshuber group:
- Purification, biophysical characterization and structure determiation of enzymes required for disulfide bond formation in the periplasm of Gram-negative bacteria.
- Mechanistic studies on the assembly of type 1 pili from pathogenic Escherichia coli strains. In vitro reconstitution of pilus assembly from all purified components. Characterization of folding, stability and assembly behaviour of individual pilus subunits.
- Identification of intermediates in the aggregation of the human Abeta peptide

Experimental work on these projects involves
- Molecular cloning, recombinant protein production in E. coli and protein purification
- Protein crystallization
- Thermodynamic and kinetic characterization of conformational changes in proteins and protein-ligand interactions by fluorescence and circular dischoism spectroscopy
- Analysis of rapid reactions by stopped-flow fluorescence
- Negative-stain electron microscopy
- Light scattering



Projects of the Weber-Ban group:

- Generation and purification of site-directed variants of the E. coli ClpA/P protease and chaperone-proteasome complexes from other organisms, their biophysical characterization, including rapid kinetics by stopped-flow methods, ATPase activity measurtements, negative-stain electron microscopy and light scattering
Lecture notesNo script
LiteratureLiterature related to the individual projects will be provided on the first day of the course.
Prerequisites / NoticeAttendance of the concept course "Biomolecular Structure and Mechanism I: Protein Structure and Function" (551-0307-00L) in the autumn semester is highly recommended for acquiring the theoretical background to this block course.