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

NEUROPHARMACOLOGY

卷:98

Review

Barden, Adam O.¹  Goler, Adam S.¹  Humphreys, Sara C.¹  Tabatabaei, Samaneh¹  Lochner, Martin²  Ruepp, Marc-David²  Jack, Thomas²  Simonin, Jonathan²  Thompson, Andrew J.³  Jones, Jeffrey P.¹  Brozik, James A.¹ 

[1] Washington State Univ, Dept Chem, Pullman, WA 99164 USA

[2] Univ Bern, Dept Chem & Biochem, CH-3012 Bern, Switzerland

[3] Univ Cambridge, Dept Pharmacol, Cambridge CB2 1PD, England

关键词: Single molecule fluorescence imaging;    Protein tracking;    P2X1;    5HT3;    Fluorescence;    LGIC;    Ligand-gated ion channel;    GPCR G-protein coupled receptor;    Molecular machine;    Stochastic;   

DOI  :  10.1016/j.neuropharm.2015.05.003

来源: Elsevier

【 摘 要 】

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. This article is part of the Special Issue entitled 'Fluorescent Tools in Neuropharmacology'. (C) 2015 Elsevier Ltd. All rights reserved.

【 授权许可】

Free