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

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