Structure and Protein-Protein Interactions of Ice Nucleation Proteins Drive Their Activity

Hartmann, S.; Ling, M.; Dreyer, L. S. A.; Zipori, A.; Finster, K. W.; Grawe, S.; Jensen, L. Z.; Borck, S.; Reicher, N.; Drace, T.; Niedermeier, D.; Jones, N. C.; Hoffmann, S. V.; Wex, H.; Rudich, Y.; Boesen, T.; Santl-Temkiv, T.

doi:10.1101/2022.01.21.477219

bioRxiv · 10.1101/2022.01.21.477219

Structure and Protein-Protein Interactions of Ice Nucleation Proteins Drive Their Activity

Hartmann, S.·Ling, M.·Dreyer, L. S. A.·Zipori, A.·Finster, K. W.·Grawe, S.·Jensen, L. Z.·Borck, S.·Reicher, N.·Drace, T.·Niedermeier, D.·Jones, N. C.·Hoffmann, S. V.·Wex, H.·Rudich, Y.·Boesen, T.·Santl-Temkiv, T.

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Abstract

ABSTRACTMicrobially-produced ice nucleating proteins (INpro) are unique molecular structures with the highest known catalytic efficiency for ice formation. Their critical role in rain formation and frost damage of crops together with their diverse commercial applications warrant an in-depth under-standing of their inherent ice nucleation mechanism. We used the machine-learning based software Al-phaFold to develop the first ab initio structural model of a bacterial INpro which is a novel beta-helix structure consisting of repeated stacks of two beta strands connected by two sharp turns. Using the synchrotron radiation circular dichroism, we validated the {beta}-strand content of the model. Combining functional studies of purified recombinant INpro, electron microscopy and modeling, we further demonstrate that the formation of dimers and higher-order oligomers is key to INpro activity. This work presents a major advance in understanding the molecular foundation for bacterial ice-nucleation activity and the basis for investigating the mechanistic role of INpro-induced ice formation in the atmosphere, and for commercial design and production of ice-nucleating particles for industrial applications.

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Hartmann, S., Ling, M., Dreyer, L. S. A., Zipori, A., Finster, K. W., Grawe, S., Jensen, L. Z., Borck, S., Reicher, N., Drace, T., Niedermeier, D., Jones, N. C., Hoffmann, S. V., Wex, H., Rudich, Y., Boesen, T., Santl-Temkiv, T.. 2022-01-22. Structure and Protein-Protein Interactions of Ice Nucleation Proteins Drive Their Activity. https://doi.org/10.1101/2022.01.21.477219

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