UV-crosslink is widely used to bind together proteins and RNA for co-isolation. Does it works also to connect proteins with other proteins? Is it possible to "reverse" such crosslink? For example, when using PFA, it is possible to "reverse" the crosslink by incubation of the sample at 70 C for 45 min.
Yes it works. Presumably it's largely driven by tyrosine dimerization but there's little research on how it works. It's quite inefficient compared to RNA but colleagues and I have done significant work crosslinking for 20 min to an hour and it works pretty well only for certain proteins. It's a caveat that should be considered when interpreting RNA crosslinking data. It shouldn't be reversible. Can be enhanced by spiking in ruthenium compounds. So far I and colleagues have crosslinked protein complexes containing FUS, TDP-43, Prc2, and telomerase in the absence of RNA. I don't think the telomerase was published and can't remember if Prc2 was. Telomerase was unusually efficient, crosslinked in minutes, and was easily seen on a gel. in below see figure 3D.
Article RNA seeds higher-order assembly of FUS protein
Yes it works well see our earlier paper.
Article Clusters of PE and PPE genes of Mycobacterium tuberculosis a...
yes it works and it can be extremely efficient with certain proteins. There is paper published last year that answers your question " Femtosecond UV-laser pulses to unveil protein–protein interactions in living cells "
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Biology
- Biomolecules
- Nucleic Acids
- RNA