In-Cell Biochemistry Using NMR Spectroscopy
Author Information
Author(s): David S. Burz, Alexander Shekhtman
Primary Institution: State University of New York at Albany
Hypothesis
How do post-translational modifications affect protein-protein interactions in living cells?
Conclusion
The study demonstrates that post-translational modifications, such as phosphorylation, can significantly alter the interaction surfaces of proteins involved in endocytosis.
Supporting Evidence
- Ubiquitin binding mediates the processivity of a large network of interactions required for proper functioning of the RTK sorting machinery.
- The methodology can be applied to any stable target molecule and may include other post-translational modifications.
- Phosphorylation of STAM2 and Hrs alters their interaction with Ubiquitin.
Takeaway
Scientists found a way to see how proteins interact inside living cells, and they learned that adding special tags to proteins can change how they connect with each other.
Methodology
The study used in-cell NMR spectroscopy to observe structural changes in protein interactions due to post-translational modifications.
Limitations
The precision of binding affinities in-cell is lower than in vitro due to variable protein expression levels.
Digital Object Identifier (DOI)
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