Tracking individual membrane proteins and their biochemistry: The power of direct observation

The advent of single molecule fluorescence microscopy has allowed experimental molecular biophysics and biochemistry to transcend traditional ensemble measurements, where the behavior of individual proteins could not be precisely sampled. The recent explosion in popularity of new super-resolution and super-localization techniques coupled with technical advances in optical designs and fast highly sensitive cameras with single photon sensitivity and millisecond time resolution have made it possible to track key motions, reactions, and interactions of individual proteins with high temporal resolution and spatial resolution well beyond the diffraction limit. Within the purview of membrane proteins and ligand gated ion channels (LGICs), these outstanding advances in single molecule microscopy allow for the direct observation of discrete biochemical states and their fluctuation dynamics. Such observations are fundamentally important for understanding molecular-level mechanisms governing these systems. Examples reviewed here include the effects of allostery on the stoichiometry of ligand binding in the presence of fluorescent ligands; the observation of subdomain partitioning of membrane proteins due to microenvironment effects; and the use of single particle tracking experiments to elucidate characteristics of membrane protein diffusion and the direct measurement of thermodynamic properties, which govern the free energy landscape of protein dimerization. The review of such characteristic topics represents a snapshot of efforts to push the boundaries of fluorescence microscopy of membrane proteins to the absolute limit.

Fair Access to Energy Resources, Market Transfers and Climate Change in the WTO

This paper addresses the issues of dual pricing and export restrictions in the energy sector, stressing the comparability of their economic and climate change impacts. It assesses whether WTO disciplines relevant and applicable to such practices are well-equipped to ensure fair access to energy resources. It finds that relevant GATT disciplines are overall deficient in the case of dual pricing and export taxes, while the landscape of WTO-plus obligations generally consisting of a network of narrowly tailored commitments. It discusses possible avenues to address such practices under the ASCM to the extent that they distort domestic energy prices and subsidize consumption of cheap fossil fuels