Journal
CELL CHEMICAL BIOLOGY
Volume 26, Issue 1, Pages 48-+Publisher
CELL PRESS
DOI: 10.1016/j.chembiol.2018.10.007
Keywords
-
Categories
Funding
- Deutsche Forschungsgemeinschaft (DFG) [SFB 1035]
- European Research Council (ERC)
- European Union [725085]
- TUM International Graduate School of Science and Engineering (IGSSE)
- European Research Council (ERC) [725085] Funding Source: European Research Council (ERC)
Ask authors/readers for more resources
Detection of dynamic protein-protein interactions within complexes and networks remains a challenging task. Here, we show by the example of the proteolytic ClpXP complex the utility of combined chemical cross-linking and mass spectrometry (XL-MS) to map interactions within ClpP and ClpX as well as across the enigmatic ClpX hexamer-ClpP heptamer interface. A few hot-spot lysines located in signature loops in ClpX were shown to be in proximity to several structural regions of ClpP providing an initial draft of the ClpX-ClpP interaction. Application of XL-MSfurther confirmed that Listeria monocytogenes ClpX interacts with the heterooligomeric ClpP1/2 complex solely via the ClpP2 apical site. Moreover, cellular interaction networks of human and bacterial proteases were elucidated via in situ chemical cross-linking followed by an antibody-based pull-down against ClpP. A subsequent mass spectro-metric analysis demonstrated an up to 3-fold higher coverage compared with co-immunoprecipitation without cross-linker revealing unprecedented insight into intracellular ClpXP networks.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available