Water-organizing Motif Continuity is Critical for Potent Ice Nucleation Protein Activity

Overview

Journal Nat Commun

Specialty Biology

Date 2022 Aug 26

PMID 36028506

Authors

Jordan Forbes

Akalabya Bissoyi

Lukas Eickhoff

Naama Reicher

Thomas Hansen

Christopher G Bon

Virginia K Walker

Thomas Koop

Yinon Rudich

Ido Braslavsky

Peter L Davies

Affiliations

Soon will be listed here.

Abstract

Bacterial ice nucleation proteins (INPs) can cause frost damage to plants by nucleating ice formation at high sub-zero temperatures. Modeling of Pseudomonas borealis INP by AlphaFold suggests that the central domain of 65 tandem sixteen-residue repeats forms a beta-solenoid with arrays of outward-pointing threonines and tyrosines, which may organize water molecules into an ice-like pattern. Here we report that mutating some of these residues in a central segment of P. borealis INP, expressed in Escherichia coli, decreases ice nucleation activity more than the section's deletion. Insertion of a bulky domain has the same effect, indicating that the continuity of the water-organizing repeats is critical for optimal activity. The ~10 C-terminal coils differ from the other 55 coils in being more basic and lacking water-organizing motifs; deletion of this region eliminates INP activity. We show through sequence modifications how arrays of conserved motifs form the large ice-nucleating surface required for potency.

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