Pannexin-3 stabilizes the transcription factor Bcl6 in a channel-independent manner to protect against vascular oxidative stress.

Targeted degradation regulates the activity of the transcriptional repressor Bcl6 and its ability to suppress oxidative stress and inflammation. Here, we report that abundance of endothelial Bcl6 is determined by its interaction with Golgi-localized pannexin 3 (Panx3) and that Bcl6 transcriptional activity protects against vascular oxidative stress. Consistent with data from obese, hypertensive humans, mice with an endothelial cell-specific deficiency in Panx3 had spontaneous systemic hypertension without obvious changes in channel function, as assessed by Ca2+ handling, ATP amounts, or Golgi luminal pH. Panx3 bound to Bcl6, and its absence reduced Bcl6 protein abundance, suggesting that the interaction with Panx3 stabilized Bcl6 by preventing its degradation. Panx3 deficiency was associated with increased expression of the gene encoding the H2O2-producing enzyme Nox4, which is normally repressed by Bcl6, resulting in H2O2-induced oxidative damage in the vasculature. Catalase rescued impaired vasodilation in mice lacking endothelial Panx3. Administration of a newly developed peptide to inhibit the Panx3-Bcl6 interaction recapitulated the increase in Nox4 expression and in blood pressure seen in mice with endothelial Panx3 deficiency. Panx3-Bcl6-Nox4 dysregulation occurred in obesity-related hypertension, but not when hypertension was induced in the absence of obesity. Our findings provide insight into a channel-independent role of Panx3 wherein its interaction with Bcl6 determines vascular oxidative state, particularly under the adverse conditions of obesity.

Solution NMR investigations of integral membrane proteins: Challenges and innovations.

Compared to soluble protein counterparts, the understanding of membrane protein stability, solvent interactions, and function are not as well understood. Recent advancements in labeling, expression, and stabilization of membrane proteins have enabled solution nuclear magnetic resonance spectroscopy to investigate membrane protein conformational states, ligand binding, lipid interactions, stability, and folding. This review highlights these advancements and new understandings and provides examples of recent applications.

Sponge-phase Lipid Droplets as Synthetic Organelles: An Ultrafast Study of Hydrogen Bonding and Interfacial Environments.

Bottom-up design of biomimetic organelles has gained recent attention as a route towards understanding the transition between non-living matter and life. Despite various artificial lipid membranes being developed, the specific relations between lipid structure, composition, interfacial properties, and morphology are not currently understood. Sponge-phase droplets contain dense, nonlamellar lipid bilayer networks that capture the complexities of the endoplasmic reticulum (ER), making them ideal artificial models of such organelles. Here, we combine ultrafast two-dimensional infrared (2D IR) spectroscopy and molecular dynamics simulations to investigate the interfacial H-bond networks in sponge-phase droplets composed of glycolipid and nonionic detergents. In the sponge phase, the interfacial environments are more hydrated and water molecules confined to the nanometer-scale aqueous channels in the sponge phase exhibit dynamics that are significantly slower compared to bulk water. Surfactant configurations and microscopic phase separation play a dominant role in determining membrane curvature and slow dynamics observed in the sponge phase. The studies suggest that H-bond networks within the nanometer-scale channels are disrupted not only by confinement but also by the interactions of surfactants, which extend 1-2 nm from the bilayer surface. The results provide a molecular-level description for controlling phase and morphology in the design of synthetic lipid organelles.

Bursting the bubble: A molecular understanding of surfactant-water interfaces.

Surfactant science has historically emphasized bulk, thermodynamic measurements to understand the microemulsion properties of greatest industrial significance, such as interfacial tensions, phase behavior, and thermal stability. Recently, interest in the molecular properties of surfactants has grown among the physical chemistry community. This has led to the application of cutting-edge spectroscopic methods and advanced simulations to understand the specific interactions that give rise to the previously studied bulk characteristics. In this Perspective, we catalog key findings that describe the surfactant-oil and surfactant-water interfaces in molecular detail. We emphasize the role of ultrafast spectroscopic methods, including two-dimensional infrared spectroscopy and sum-frequency-generation spectroscopy, in conjunction with molecular dynamics simulations, and the role these techniques have played in advancing our understanding of interfacial properties in surfactant microemulsions.

Ions Slow Water Dynamics at Nonionic Surfactant Interfaces.

Nonionic surfactant interfaces are often considered to be unaffected by aqueous ions. However, ions often localize to interfaces and, as such, can interact with surfactants either via direct contact or by affecting interfacial hydrogen bond structures in water. Characterizing these effects is essential to understanding how ions affect interfacial properties at the oil-water interface in the presence of nonionic surfactants. Here, we use two-dimensional infrared (2D IR) spectroscopy in combination with atomistic molecular dynamics (MD) simulations to study the effects of high-concentration Na+ and Ca2+ ions on interfacial hydrogen bond dynamics in heterogeneous sorbitan stearate reverse micelles. Experiments show only minor changes in interfacial hydrogen bond populations when salts are added but those interfacial water network dynamics are slowed by nearly 300%. Molecular dynamics simulations show the slowdown results from an increased disorder in surfactant headgroup orientation and packing density, which stabilizes hydrogen bonding interactions between surfactants and interfacial water.

Slow Oil, Slow Water: Long-Range Dynamic Coupling across a Liquid-Liquid Interface.

Liquid interfaces are dynamic environments characterized by chemical and physical properties that are different from the bulk. Here we use ultrafast, two-dimensional infrared spectroscopy and molecular dynamics simulations to measure the effect of oil phase viscosity on interfacial water dynamics in reverse micelles. The oil and water phases are separated by a 2.3-3.0 nm surfactant interface. Increasing the oil viscosity from 0.3 to 10.5 cP slows down water motions from 0.9 to 1.5 ps, as measured by the carbonyl frequency fluctuations. Simulations, which agree semi-quantitatively with experiments, show that water self-diffusion at the interface slows with increased oil viscosity. This study shows that the oil phase "transmits" its dynamics to the aqueous phase through the surfactant layer with minimal perturbations to surfactant-water interfacial structure.

Interfacial H-Bond Dynamics in Reverse Micelles: The Role of Surfactant Heterogeneity.

Characterizing the hydrogen bond structure and dynamics at surfactant interfaces is essential for understanding how microscopic interactions translate to bulk microemulsion properties. Heterogeneous blends containing tens or hundreds of surfactants are common in the industry, but the most fundamental studies have been carried out on micelles composed of a single surfactant species. Therefore, the effect of surfactant heterogeneity on the interfacial structure and dynamics remains poorly understood. Here, we use ultrafast two-dimensional infrared spectroscopy and molecular dynamics simulations to characterize sub-picosecond solvation dynamics as a function of the surfactant composition in ∼120 nm water-in-oil reverse micelles. We probe the ester carbonyl vibrations of nonionic sorbitan surfactants, which are located precisely at the interface between the polar and nonpolar regions, and as such, report on the interfacial water dynamics. We show a 7% increase in hydrogen bond populations together with a 37% slowdown of interfacial hydrogen bond dynamics in heterogeneous mixtures containing hundreds of species, compared to more uniform compositions. Simulations, which are in semiquantitative agreement with experiments, indicate that structural diversity leads to decreased packing efficiency, which in turn drives water further into the otherwise hydrophobic region. Interestingly, this increase in hydration is accompanied by a slowdown of dynamics, indicating that water molecules solvating surfactants are conformationally constrained. These studies demonstrate that the composition and heterogeneity are key factors in determining interfacial properties.