Ice Recrystallization Unveils the Binding Mechanism Operating at a Diffused Interface.

Recrystallization of ice is a natural phenomenon that causes adverse effects in cryopreservation, agriculture, and in frozen food industry. It has long been recognized that ice recrystallization occurs through the Ostwald ripening and accretion processes. However, neither of these processes has been explored in microscopic detail by state-of-the-art experimental techniques. We carried out atomistic molecular dynamics (MD) simulations to explore ice recrystallization through the accretion process. Attempts have been made to elucidate the binding mechanism that is operating at the diffused ice-water interface. It is demonstrated that two ice crystals spontaneously recognize each other and bind together to form a large crystal in liquid water, resulting in ice recrystallization by accretion. Interestingly, the study reveals that the binding occurs due to the freezing of the interfacial water layer present between the two ice planes, even at a temperature above the melting point of the ice crystal. The synergistically enhanced ordering effect of two ice surfaces on the interfacial water leads to such freezing occurring during the binding process. However, proper crystallographic alignment is not necessarily required for the binding of the two crystals. Simulations have also been carried out to study the binding between an ice crystal and the model ice-binding surface (IBS) of an antifreeze protein above the melting point of the ice crystal. It is found that such binding at the IBS is accompanied by freezing of the interfacial water. This establishes that the synergetic ordering-driven freezing of interfacial water is a common binding mechanism at the diffused surfaces of ice crystals. We believe that this mechanism will provide a microscopic understanding of the process of recrystallization inhibition and thus help in designing suitable materials for potent applications in recrystallization inhibition.

Elucidating the Sluggish Water Dynamics at the Ice-Binding Surface of the Hyperactive Tenebrio molitor Antifreeze Protein.

Quasi-ice-like hydration waters on the ice-binding surface (IBS) of an antifreeze protein (AFP) commonly exhibit sluggish dynamics especially at low temperatures. In this work, we have analyzed molecular dynamics (MD) simulation trajectories at two different temperatures for Tenebrio molitor antifreeze protein (TmAFP) to explore whether the unique quasi-ice-like structuring of hydration water has any impact on making their dynamics slower on the IBS of the protein. Our calculation reveals that, as translational dynamics is coupled with the conformational fluctuations, hydration water on the IBS exhibits sluggish translational motion due to reduced flexibility of the IBS compared to that on the non-ice-binding surface (NIBS) of the protein. Interestingly, it is noticed that rotational motion of hydration water is not coupled with the conformational fluctuations of the surfaces. In that case, structural relaxations of the protein-water (PW) and water-water (WW) hydrogen bonds compete with each other to make the rotational dynamics of hydration water around the IBS either faster or slower with respect to those around the NIBS. At low temperature, the slower structural relaxation of water-water hydrogen bonds dominates and imparts sluggish rotational motion of the hydration water on the IBS of the protein. The slower structural relaxation of water-water hydrogen bonds and hence the retarded rotational dynamics, despite the weak short-lived PW hydrogen bonds on the IBS, is clearly a manifestation of the rigid quasi-ice-like structure of the hydration shell on that surface.

Role of Polar and Nonpolar Groups in the Activity of Antifreeze Proteins: A Molecular Dynamics Simulation Study.

Molecular dynamics simulations have been carried out separately with the hyperactive Tenebrio molitor antifreeze protein ( TmAFP) and with its nonactive mutant at 300 K to elucidate the role of polar and nonpolar groups in the activities of antifreeze proteins (AFPs). Simulation results reveal that both polar and nonpolar groups contribute to develop the required quasi-ice-like hydration layer on the ice-binding surface (IBS) of an AFP for binding onto ice. Nonpolar groups on the IBS induce the formation of locally ordered icelike low-density waters in the hydration layer through hydrophobic interactions, and polar groups of the surface integrate these waters into a quasi-ice-like layered structure through hydrogen-bonding interactions. These contributions of polar and nonpolar groups apparently contradict the behavior of winter flounder antifreeze protein (wfAFP) mutants possibly due to switching of IBS of wfAFP upon mutation of threonine residues with valine residues.

Operation of Kelvin Effect in the Activities of an Antifreeze Protein: A Molecular Dynamics Simulation Study.

Ice growth and melting inhibition activities of antifreeze proteins (AFPs) are better explained by the adsorption-inhibition mechanism. Inhibition occurs as a result of the Kelvin effect induced by adsorbed protein molecules onto the surface of seed ice crystal. However, the Kelvin effect has not been explored by the state-of-the-art experimental techniques. In this work, atomistic molecular dynamics simulations have been carried out with Tenebrio molitor antifreeze protein ( TmAFP) placed at ice-water interface to probe the Kelvin effect in the mechanism of AFPs. Simulations show that, below equilibrium melting temperature, ice growth is inhibited through the convex ice-water interface formation toward the water phase and, above equilibrium melting temperature, ice melting is inhibited through the concave ice-water interface formation inward to ice phase. Simulations further reveal that the radius of curvature of the interface formed to stop the ice growth increases with decrease in the degree of supercooling. Our results are in qualitative agreement with the theoretical prediction of the Kelvin effect and thus reveal its operation in the activities of AFPs.

Interfacial Water Arrangement in the Ice-Bound State of an Antifreeze Protein: A Molecular Dynamics Simulation Study.

Molecular dynamics (MD) simulations have been carried out to study the heterogeneous ice nucleation on modeled peptide surfaces. Simulations show that large peptide surfaces made by TxT (threonine-x-threonine) motifs with the arrangements of threonine (Thr) residues identical to the periodic arrangements of waters on either the basal or prism plane of ice are capable of ice nucleation. Nucleated ice plane is the (0001) basal plane of hexagonal ice (Ih) or (111) plane of cubic ice (Ic). However, due to predefined simulation cell dimensions, the ice growth is only observed on the surface where the Thr residues are arranged like the water arrangement on the basal plane of ice Ih. The γ-methyl and γ-hydroxyl groups of Thr residue are necessary for such ice formation. From this ice nucleation and growth simulation, the interfacial water arrangement in the ice-bound state of Tenebrio molitor antifreeze protein (TmAFP) has been determined. The interfacial water arrangement in the ice-bound state of TmAFP is characterized by five-membered hydrogen bonded rings, where each of the hydroxyl groups of the Thr residues on the ice-binding surface (IBS) of the protein is a ring member. It is found that the water arrangement at the protein-ice interface is distorted from that in bulk ice. Our analysis further reveals that the hydroxyl groups of Thr residues on the IBS of TmAFP form maximum three hydrogen bonds each with the waters in the bound state and methyl groups of Thr residues occupy wider spaces than the normal grooves on the (111) plane of ice Ic. Methyl groups are also located above and along the 3-fold rotational axes of the chair-formed hexagonal hydrogen bonded water rings on the (111) plane.

Hydration behavior at the ice-binding surface of the Tenebrio molitor antifreeze protein.

Molecular dynamics (MD) simulations have been carried out at two different temperatures (300 and 220 K) to study the conformational rigidity of the hyperactive Tenebrio molitor antifreeze protein (TmAFP) in aqueous medium and the structural arrangements of water molecules hydrating its surface. It is found that irrespective of the temperature the ice-binding surface (IBS) of the protein is relatively more rigid than its nonice-binding surface (NIBS). The presence of a set of regularly arranged internally bound water molecules is found to play an important role in maintaining the flat rigid nature of the IBS. Importantly, the calculations reveal that the strategically located hydroxyl oxygens of the threonine (Thr) residues in the IBS influence the arrangements of five sets of ordered waters around it on two parallel planes that closely resemble the basal plane of ice. As a result, these waters can register well with the ice basal plane, thereby allowing the IBS to preferentially bind at the ice interface and inhibit its growth. This provides a possible molecular reason behind the ice-binding activity of TmAFP at the basal plane of ice.