Structural thermodynamics by cryo-EM resolves minute energy differences in heterogeneous protein complexes.
Boyens-Thiele, L.; Bicanin, M.; Engholm-Keller, K.; Boggild, A.; Boesen, T.; Svensson, B.; Buell, A. K.; Morth, J. P.
Show abstract
Hetero-oligomeric protein complexes play crucial roles across biology, providing combinatorial diversity and context-specific functionality in membrane receptors, molecular chaperones and beyond. Yet, existing methods are limited in their ability to resolve and quantify the assembly energetics of complex biomolecular populations. Here, we introduce a comprehensive method combining iterative 3D classification, error-rate quantification via simulated particles, and global thermodynamic modeling to extract interaction energies from cryo-EM data. Using hexameric cruciferin (the primary rapeseed storage protein) as a model, we reconstruct a 1.83 [A] structure and distinguish three highly similar 50 kDa isoforms. We resolve the complex compositional heterogeneity of the hexameric species at the single-particle level and fit a global binding model revealing pairwise interaction energies between isoforms differing by fractions of the thermal energy (kB T ). This method offers a powerful, broadly applicable tool to characterize the compositional heterogeneity and energetics of biomolecular complexes.
Matching journals
The top 4 journals account for 50% of the predicted probability mass.
Similar papers in this journal
- Integrated NMR and cryo-EM atomic-resolution structure determination of a half-megadalton enzyme complex 95%
- Cryo-EM Structures of Higher Order Gephyrin OligomersReveal Principles of Inhibitory Postsynaptic Scaffold Organization 95%
- Structural basis of Cullin-2 RING E3 ligase regulation by the COP9 signalosome 94%
Similar papers in this journal
Similar papers in this journal
- Absolute hand determination of glycofibrils from natural sources in cryo-EM 95%
- Capturing ribosomal structures in cellular extracts with cryoPRISM: A purification-free cryoEM approach reveals novel structural states 95%
- Microfluidic protein isolation and sample preparation for high resolution cryo-EM 95%
Similar papers in this journal
Similar papers in this journal
"Similar papers" are the closest papers from that journal in the model's embedding space. They show what the match is built on, but the ranking comes mostly from a classifier over the whole training set, not from these examples alone.