Inventor(s): Shail N. Shah (Ann Arbor, MI), Shinji Oita (Ann Arbor, MI) Assignee(s): Toyota Motor Engineering Manufacturing North America, Inc. (Plano, TX) Law Firm: Darrow Mustafa PC (2 non-local offices) Application No., Date, Speed: 15219424 on 07/26/2016 (868 days app to issue)

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Inventor(s): Timothy J. Bennett (Coloardo Springs, CO) Assignee(s): TUESDAY MORNING PARTNERS, LTD. (Dallas, TX) Law Firm: Haynes and Boone, LLP (Local + 13 other metros) Application No., Date, Speed: 29549460 on 12/22/2015 (1085 days app to issue)

The present study provides the first observation of chiral dipole orientated water wires within artificial water channels under ambient bilayer membrane conditions. The dipolar water alignment within the membrane is driven by the orientation and ordering of imidazole/water dipoles. These results demonstrate that the water dipolar orientation is preserved from the single-crystal structures to the inherently dynamical water wires confined in artificial water channels in bilayer membranes. Overall, this system parallels the world of water transport through natural proteins, where the picture of dipolar water structures confined within a pore is the basis of the high permeability/selectivity of AQPs (24). Our results underline the chirality translation from lipids to I-quartet channels and from channels to structured dipolar water wires. First, we observe from the x-ray single-crystal structures that induced chirality in the channels and the structured water in the channel pores adapt accordingly. Here, the chiral backbone of the chiral S-HC8 channels induces a unique chiral dipolar orientation for water wires along the axis of the pores as compared with the achiral HC8 channels that present alternating chiral dipole orientations of water wires. This architecture further translated into a performance difference when assessing water permeability: the net permeabilities of HC8 and its chiral isomers R-HC8 and S-HC8 at a channel-to-lipid weight ratio of 1 were 1.0 ± 0.3 μm/s, 2.8 ± 0.7 μm/s, and 4.1 ± 0.2 μm/s, as previously determined (14). The MD simulations have further shown that these subtle differences in structure can induce energetic differences when considering the strength of the hydrogen bonding network in the confinement region of the channels. The well-structured water wires in the S-HC8 channels have an almost unperturbed behavior, with water flowing freely without significant energy penalties. This observation is mainly due to the fact that water found in the pores has 3.2 to 3.4 hydrogen bonds, which are very similar to the hydrogen bonds that occur in bulk water. On the other hand, HC8 offers a rather less favorable confinement and stabilization for the water wires. The strong water dipole alignment inside the channels is confirmed by the SFG results, where the signal intensity is higher for both chiral and achiral dipolar water wires, as well as strongly bound water molecules in the case of the S-HC8 embedded SLBs compared with the reference SLB. The SLBs are supported on a CaF2 prism coated with a 100-nm layer of SiO2, produce an asymmetry in the entire system, and induce a weak chiral alignment in the interfacial water structure through the chiral constituent lipids. The confined water wires in both the HC8 and the S-HC8 channels furthermore exhibit a strong chiral ordering template by the chiral channel structure.

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