Chemical and site-directed cross-linking allows the analysis of interactions between proteins and/or their subunits. Therefore, it is highly relevant for the SFB 1381 “Dynamic Organization of Cellular Protein Machineries”.
This workshop, organized by our B06 project in collaboration with Z01, will give an overview on the concept of chemical and site-directed cross-linking. Various cross-linking methods and cross-linkers will be introduced, followed by a detailed discussion of their respective advantages and limitations. It will be demonstrated how cross-linkers as well as the residues for site-directed and chemical cross-linking can be determined, e.g. using protein AlphaFold prediction. As an example, an in vivo paraformaldehyde experiment will be shown and in an interactive session a cross-linking experiment will be designed. Typical pitfalls and recommendations for adapting protocols will be debated.
Finally, the analysis of the covalently cross-linked products using biochemical and mass spectrometry-based methods will be illustrated. The objectives range from identifying unknown interaction partners to uncovering detailed structural and topological information.
Participants: PhD students and PostDocs, PIs are welcome. Experience in cross-linking is not required; however, general biochemical lab skills are assumed.
Date: March 27/28 2025 (in total 1.5 days)
Location: ZBMZ, Stefan-Meier-Str. 17, large seminar room, 1st floor, Room Nr 01 006
If you are interested in participation, please contact the SFB coordination office (sfb1381@biochemie.uni-freiburg.de).
