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High-speed cryo-microscopy reveals that ice-nucleating proteins of Pseudomonas syringae trigger freezing at hydrophobic interfaces

2024/07/03 by Paul Bieber, Nadine Borduas‐Dedekind · 1 voice
Earth and Planetary Sciences · Environmental Science · Biochemistry, Genetics and Molecular Biology · #nanoparticles nucleation surface interactions #Physiological and biochemical adaptations #Insect and Arachnid Ecology and Behavior

paper · pdf · doi:10.1126/sciadv.adn6606

openalex publication_date 2024/07/03 · openalex created_date 2024/07/06 · openalex updated_date 2026/06/13

Abstract

Ice-nucleating proteins (INpro) trigger the freezing of supercooled water droplets relevant to atmospheric, biological, and technological applications. The high ice nucleation activity of INpro isolated from the bacteria Pseudomonas syringae could be linked to the aggregation of proteins at the bacterial membrane or at the air-water interface (AWI) of droplets. Here, we imaged freezing onsets, providing direct evidence of these proposed mechanisms. High-speed cryo-microscopy identified the onset location of freezing in droplets between two protein-repellent glass slides. INpro from sterilized P. syringae (Snomax) statistically favored nucleation at the AWI of the droplets. Removing cellular fragments by filtration or adding surfactants increased the frequency of nucleation events at the AWI. On the other hand, cultivated intact bacteria cells or lipid-free droplets nucleated ice without an affinity to the AWI. Overall, we provide visual evidence that INpro from P. syringae trigger freezing at hydrophobic interfaces, such as the AWI or the bacterial membrane, with important mechanistic implications for applications of INpro.

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