Protein folding

We are examining protein folding and unfolding characteristics by the use of fluorescent markers.
In collaboration with Prof. Hartl, we have examined in detail the functionality of chaperonin-assisted protein folding. The GroEL/GroES system from E. coli works as a container (GroEL) with a lid (GroES). Maltose binding protein (MBP) was chosen as a substrate and labeled with two fluorescent dyes at carefully selected labeling positions. In spFRET experiments the folding state of MBP and the influence of the chaperonin-system were observed by monitoring the FRET efficiency. Different mutants of the GroEL/GroES system were examined to clarify its working mechanism.
GroEl action
In another project, a collaboration with the group of Prof. Buchner, we study the folding and unfolding pathways of different immunoglobulin (Ig) antibody domains and their interaction with each other. By labeling a protein with two different fluorescent dyes at different labeling positions, the dyes will undergo FRET. The FRET efficiency E depends on the spatial distance of the dyes, thus indicating the folding state of the protein. At the same time, a third fluorescent marker attached to a different Ig-domain enables us to determine the colocalization and thereby the fusion of the domains using PIE-FCCS.

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