Protein Quaternary Structures in Solution are a Mixture of Multiple forms

Marciano, S.; Debabrata, D.; Listov, D.; Fleishman, S. J.; Sonn-Segev, A.; Mertens, H.; Busch, F.; Kim, Y.; Harvey, S. R.; Wysocki, V. H.; Schreiber, G.

doi:10.1101/2022.03.30.486392

bioRxiv · 10.1101/2022.03.30.486392

Protein Quaternary Structures in Solution are a Mixture of Multiple forms

Marciano, S.·Debabrata, D.·Listov, D.·Fleishman, S. J.·Sonn-Segev, A.·Mertens, H.·Busch, F.·Kim, Y.·Harvey, S. R.·Wysocki, V. H.·Schreiber, G.

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Abstract

Over half the proteins form homo or hetero-oligomeric structures. Experimentally determined structures are often considered in determining a proteins oligomeric state, but static structures miss the dynamic equilibrium between different quaternary forms. The problem is exacerbated in homo-oligomers, where the oligomeric states are challenging to characterize. Here, we re-evaluated the oligomeric state of 17 different bacterial proteins across a broad range of protein concentrations and solutions by native mass-spectrometry (MS), mass photometry (MP), size exclusion chromatography (SEC), and small-angle x-ray scattering (SAXS), finding that most exhibit several oligomeric states. Surprisingly, many proteins did not show mass-action driven equilibrium between the oligomeric states. For approximately half the proteins, the predicted oligomeric forms described in publicly available databases underestimated the complexity of protein quaternary structures in solution. Conversely, AlphaFold Multimer provided an accurate description of the potential multimeric states for most proteins, suggesting that it could help resolve uncertainties on the solution state of many proteins.

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Marciano, S., Debabrata, D., Listov, D., Fleishman, S. J., Sonn-Segev, A., Mertens, H., Busch, F., Kim, Y., Harvey, S. R., Wysocki, V. H., Schreiber, G.. 2022-03-31. Protein Quaternary Structures in Solution are a Mixture of Multiple forms. https://doi.org/10.1101/2022.03.30.486392

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