Characterizing oligomerization dynamics of an E3 ligase using mass photometry

In this webinar, Refeyn travel grant winner Xiaojing Huang explains how her research harnessed the power of mass photometry to analyze the oligomeric state of the DCAF15-DDA1-DDB1 E3 ligase at low nanomolar concentrations. This study unveiled, for the first time, the formation of dimers and trimers with quantified dissociation constants. The oligomeric remodeling effect emphasized the transition from trimer to monomer upon molecular glue addition, shedding light on how molecular glues regulate E3 ligase oligomerization at physiological concentration.


TIMESTAMPS

00:00 – Moderator introduces Xiaojing
02:03 – Xiaojing’s introduction and acknowledgements
02:50 – Undruggable proteins and molecular glues
05:43 – The ubiquitin-proteasome system
07:29 – Protein degraders in medicinal chemistry
11:29 – Introduction to the DCAF15-DDA1-DDB1 E3 ligase
12:58 – Native mass spectrometry for measuring protein-protein interactions
14:43 – Mass photometry for determining protein oligomeric states
15:11 – DCAF15-DDA1-DDB1 oligomers measured with native mass spectrometry
16:52 – DCAF15-DDA1-DDB1 oligomers measured with mass photometry
19:24 – Characterizing oligomerization and regulation of DCAF15-DDA1-DDB1
21:46 – The DCAF15-DDA1-DDB1, RBM39 and molecular glue ternary complex
25:05 – Measuring glue efficacy with native mass spectrometry
26:27 – Measuring glue efficacy with AlphaScreen
27:37 – Native mass spectrometry agrees with AlphaScreen
28:31 – Native mass spectrometry can quantify glue efficacy
30:46 – Weak E3 ligase-target interactions can be detected with native mass spectrometry
33:50 – Conclusions
34:58 – Future work
35:49 – Acknowledgements

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FURTHER RESOURCES:

Brochure – Biomolecular characterization with mass photometry - https://info.refeyn.com/mass-photomet...
Application note – Characterizing protein oligomerization with automated mass photometry - https://info.refeyn.com/characterizin...
Application note – Formation of PROTAC ternary complexes measured with mass photometry - https://info.refeyn.com/app-note-protacs
Webinar – Mass photometry to study the oligomerization of the ubiquitin ligase HACE1 -    • Mass photometry to study the oligomer...  

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