How Size and Aggregation of Ice-Binding Proteins Control Their Ice Nucleation Efficiency

Qiu, Yuqing; Hudait, Arpa; Molinero, Valeria

doi:10.1021/jacs.9b01854

Cited by 122 publications

(223 citation statements)

References 87 publications

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“…Oligomerization has been reported previously for other InaZ constructs and related β-helix structures of hyperactive insect anti-freeze proteins (AFPs). 19,27,44,45 In summary, we conclude that the β-helix model agrees with the IR data and describes the solution structure of InaZ9R well.…”

Section: Figure 2 Experimental and Calculated Ft-ir (Top) And Parallsupporting

confidence: 76%

“…56 Large areas of ice-nucleation active sites are crucial for heterogeneous ice nucleation. 44,56 At cell surfaces, based on current models, InaZ is anchored through its N-terminal domain 3 and would presumably be flexible enough to reorient from perpendicular to parallel to the cell surface with decreasing temperatures. Thus, a reorientation of InaZ could also explain the puzzle of why water close to ice-active bacterial cells becomes more ordered when cooled to lower temperatures.…”

Section: Resultsmentioning

confidence: 99%

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The Ice Nucleating Protein InaZ is Activated by Low Temperature

Roeters¹,

Golbek²,

Bregnhøj³

et al. 2020

Preprint

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Ice-nucleation active (INA) bacteria can promote the growth of ice more effectively than any other known material. Utilizing specialized ice-nucleating proteins (INPros), they obtain nutrients from plants by inducing frost damage and, when airborne in the atmosphere, they drive ice nucleation within clouds and may affect global precipitation patterns. Despite their evident environmental importance, the molecular mechanisms behind INProinduced freezing have remained largely elusive. In the present study, we investigated the folding and the structural basis for interactions between water and the ice-nucleating protein InaZ from the INA bacterium Pseudomonas syringae strain R10.79. Using vibrational sum-frequency generation 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, hydrogen bonding between INPros 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 spectral calculations and molecular-dynamics simulations shows that the INPro reorients at lower temperatures. We suggest that the reorientation can enhance order-inducing water interactions and, thereby, the effectiveness of ice nucleation by InaZ.

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Section: Figure 2 Experimental and Calculated Ft-ir (Top) And Parallsupporting

confidence: 76%

Section: Resultsmentioning

confidence: 99%

The Ice Nucleating Protein InaZ is Activated by Low Temperature

Roeters¹,

Golbek²,

Bregnhøj³

et al. 2020

Preprint

View full text Add to dashboard Cite

show abstract

“…Figure B shows a simulation of spruce budworm AFP and its hypothesized binding to the prism plane via coordinated water molecules providing a match . Whilst the molecular binding details are still under investigation, there is overwhelming evidence for AFPs binding to ice faces and strong evidence of molecular level interactions have been determined . However, for the facially amphipathic molecules, with no evidence for ice binding, an alternative molecular level mechanism which can give rise to IRI is required.…”

Section: Facially Amphiphilic Non‐ice‐binding Materials and Compoundsmentioning

confidence: 99%

“…[54] Whilst the molecular binding details are still under investigation, there is overwhelming evidence for AFPs binding to ice faces and strong evidence of molecular level interactions have been determined. [34,[55][56][57][58][59] However, for the facially amphipathic molecules, with no evidence for ice binding, an alternative molecular level mechanism which can give rise to IRI is required. One proposal is that these can sit at the semiordered water layer (sometimes referred to as quasi-liquid layer, QLL, which is strictly a definition at surfaces) [60] / bulk water interface, rather than directly interact with ice faces.…”

Section: Facially Amphiphilic Non-ice-binding Materials and Compoundsmentioning

confidence: 99%

Mimicking the Ice Recrystallization Activity of Biological Antifreezes. When is a New Polymer “Active”?

Biggs

Stubbs

Graham

et al. 2019

Macromolecular Bioscience

111

179

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Antifreeze proteins and ice‐binding proteins have been discovered in a diverse range of extremophiles and have the ability to modulate the growth and formation of ice crystals. Considering the importance of cryoscience across transport, biomedicine, and climate science, there is significant interest in developing synthetic macromolecular mimics of antifreeze proteins, in particular to reproduce their property of ice recrystallization inhibition (IRI). This activity is a continuum rather than an “on/off” property and there may be multiple molecular mechanisms which give rise to differences in this observable property; the limiting concentrations for ice growth vary by more than a thousand between an antifreeze glycoprotein and poly(vinyl alcohol), for example. The aim of this article is to provide a concise comparison of a range of natural and synthetic materials that are known to have IRI, thus providing a guide to see if a new synthetic mimic is active or not, including emerging materials which are comparatively weak compared to antifreeze proteins, but may have technological importance. The link between activity and the mechanisms involving either ice binding or amphiphilicity is discussed and known materials assigned into classes based on this.

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“…Structures of the investigated proteins and the tobacco mosaic virus: horse spleen (apo)ferritin cage and (apo)ferritin monomer (PDB ID: 4V1W, Russo and Passmore, 2014), chicken ovalbumin (PDB ID: 1OVA,Stein et al, 1991), hydrophobin class I DewA (PDB ID: 2LSH;Morris et al, 2013b), casein micelle consisting of the four different casein proteins: α s1 -casein (purple), α s2 -casein (light blue), β-casein (red), κ-casein (dark blue with tail) (adapted fromRebouillat and Ortega-Requena, 2015), ice-binding protein LeIBP dimer (PDB ID: 3UYU,Lee et al, 2012), and tobacco mosaic virus (TMV) (PDB ID: 3J06;Ge and Zhou, 2011).…”

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confidence: 99%

Protein aggregates nucleate ice: the example of apoferritin

et al. 2020

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Abstract. Biological material has gained increasing attention recently as a source of ice-nucleating particles that may account for cloud glaciation at moderate supercooling. While the ice-nucleation (IN) ability of some bacteria can be related to membrane-bound proteins with epitaxial fit to ice, little is known about the IN-active entities present in biological material in general. To elucidate the potential of proteins and viruses to contribute to the IN activity of biological material, we performed bulk freezing experiments with the newly developed drop freezing assay DRoplet Ice Nuclei Counter Zurich (DRINCZ), which allows the simultaneous cooling of 96 sample aliquots in a chilled ethanol bath. We performed a screening of common proteins, namely the iron storage protein ferritin and its iron-free counterpart apoferritin, the milk protein casein, the egg protein ovalbumin, two hydrophobins, and a yeast ice-binding protein, all of which revealed IN activity with active site densities > 0.1 mg−1 at −10 ∘C. The tobacco mosaic virus, a plant virus based on helically assembled proteins, also proved to be IN active with active site densities increasing from 100 mg−1 at −14 ∘C to 10 000 mg−1 at −20 ∘C. Among the screened proteins, the IN activity of horse spleen ferritin and apoferritin, which form cages of 24 co-assembled protein subunits, proved to be outstanding with active site densities > 10 mg−1 at −5 ∘C. Investigation of the pH dependence and heat resistance of the apoferritin sample confirmed the proteinaceous nature of its IN-active entities but excluded the correctly folded cage monomer as the IN-active species. A dilution series of apoferritin in water revealed two distinct freezing ranges, an upper one from −4 to −11 ∘C and a lower one from −11 to −21 ∘C. Dynamic light scattering measurements related the upper freezing range to ice-nucleating sites residing on aggregates and the lower freezing range to sites located on misfolded cage monomers or oligomers. The sites proved to persist during several freeze–thaw cycles performed with the same sample aliquots. Based on these results, IN activity seems to be a common feature of diverse proteins, irrespective of their function, but arising only rarely, most probably through defective folding or aggregation to structures that are IN active.

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How Size and Aggregation of Ice-Binding Proteins Control Their Ice Nucleation Efficiency

Cited by 122 publications

References 87 publications

The Ice Nucleating Protein InaZ is Activated by Low Temperature

The Ice Nucleating Protein InaZ is Activated by Low Temperature

Mimicking the Ice Recrystallization Activity of Biological Antifreezes. When is a New Polymer “Active”?

Protein aggregates nucleate ice: the example of apoferritin

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