Ice-nucleating proteins are activated by low temperatures to control the structure of interfacial water

Nat Commun. 2021 Feb 19;12(1):1183. doi: 10.1038/s41467-021-21349-3.


Ice-nucleation active (INA) bacteria can promote the growth of ice more effectively than any other known material. Using specialized ice-nucleating proteins (INPs), they obtain nutrients from plants by inducing frost damage and, when airborne in the atmosphere, they drive ice nucleation within clouds, which may affect global precipitation patterns. Despite their evident environmental importance, the molecular mechanisms behind INP-induced freezing have remained largely elusive. We investigate the structural basis for the interactions between water and the ice-nucleating protein InaZ from the INA bacterium Pseudomonas syringae. Using vibrational sum-frequency generation (SFG) and two-dimensional infrared spectroscopy, we demonstrate that the ice-active repeats of InaZ adopt a β-helical structure in solution and at water surfaces. In this configuration, interaction between INPs and water molecules imposes structural ordering on the adjacent water network. The observed order of water increases as the interface is cooled to temperatures close to the melting point of water. Experimental SFG data combined with molecular-dynamics simulations and spectral calculations show that InaZ reorients at lower temperatures. This reorientation can enhance water interactions, and thereby the effectiveness of ice nucleation.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Atmosphere
  • Bacterial Outer Membrane Proteins / chemistry*
  • Bacterial Outer Membrane Proteins / genetics
  • Bacterial Outer Membrane Proteins / metabolism*
  • Cold Temperature*
  • Deuterium Oxide
  • Freezing
  • Ice
  • Molecular Dynamics Simulation
  • Plants / microbiology
  • Pseudomonas syringae / metabolism
  • Water / chemistry*


  • Bacterial Outer Membrane Proteins
  • Ice
  • ice nucleation protein
  • Water
  • Deuterium Oxide

Associated data

  • figshare/10.6084/m9.figshare.13372793