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Abstract:

At the nanoscale level biological membranes present a complex interface with the solvent. The functional dynamics and relative flexibility of membrane components together with the presence of specific ionic effects can combine to create exciting new phenomena that challenge traditional theories such as the Derjaguin-Landau-Verwey-Overbeek (DLVO) theory or models interpreting the role of ions in terms of their ability to structure water (structure making/breaking). Here we investigate ionic effects at the surface of a highly charged extremophile membrane composed of a proton pump (bacteriorhodopsin) and archaeal lipids naturally assembled into a 2D crystal. Using amplitude-modulation atomic force microscopy (AM-AFM) in solution, we obtained sub-molecular resolution images of ion-induced surface restructuring of the membrane. We demonstrate the presence of a stiff cationic layer condensed at its extracellular surface. This layer cannot be explained by traditional continuum theories. Dynamic force spectroscopy experiments suggest that it is produced by electrostatic correlation mediated by a Manning-type condensation of ions. In contrast, the cytoplasmic surface is dominated by short-range repulsive hydration forces. These findings are relevant to archaeal bioenergetics and halophilic adaptation. Importantly, they present experimental evidence of a natural system that locally controls its interactions with the surrounding medium and challenges our current understanding of biological interfaces.

Abstract:

Solid-liquid interfaces play a fundamental role in surface electrochemistry, catalysis, wetting, self-assembly and biomolecular functions. The interfacial energy determines many of the properties of such interfaces, including the arrangement of the liquid molecules at the surface of the solid. Diffraction techniques are often used to investigate the structure of solid-liquid interfaces, but measurements of irregular or inhomogeneous interfaces remain challenging. Here, we report atomic-and molecular-resolution images of various organic and inorganic samples in liquids, obtained with a commercial atomic force microscope operated dynamically with small-amplitude modulation. This approach uses the structured liquid layers close to the solid to enhance lateral resolution. We propose a model to explain the mechanism dominating the image formation, and show that the energy dissipated during this process is related to the interfacial energy through a readily achievable calibration curve. Our topographic images and interfacial energy maps could provide insights into important interfaces. © 2010 Macmillan Publishers Limited. All rights reserved.

Abstract:

Membrane proteins are laterally coupled to the surrounding cell membrane through complex interactions that can modulate their function. Here, we directly observe and quantify the dynamics of functioning bacteriorhodopsin (bR) in its native membrane, a crystalline aggregate of bR trimers. We show that much of a monomer's isomerization energy is mechanically redistributed into the membrane, producing cooperative activity within the trimer while simultaneously generating functionally relevant long-range lateral pressure waves. Our results provide evidence of coordinated short and long-range effects in the cell membrane. © 2009 The Royal Society of Chemistry.