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

Barden, Adam O.; Goler, Adam S.; Humphreys, Sara C.; Tabatabaei, Samaneh; Lochner, Martin; Ruepp, Marc-David; Jack, Thomas; Simonin, Jonathan; Thompson, Andrew James; Jones, Jeffrey P.; Brozik, James A. (2015). Tracking individual membrane proteins and their biochemistry: The power of direct observation. Neuropharmacology, 98, pp. 22-30. Elsevier 10.1016/j.neuropharm.2015.05.003

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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.

Item Type:

Journal Article (Review Article)

Division/Institute:

08 Faculty of Science > Departement of Chemistry and Biochemistry

UniBE Contributor:

Lochner, Martin; Ruepp, Marc-David; Jack, Thomas; Simonin, Jonathan and Thompson, Andrew James

Subjects:

500 Science > 570 Life sciences; biology
500 Science > 540 Chemistry

ISSN:

0028-3908

Publisher:

Elsevier

Language:

English

Submitter:

Martin Lochner

Date Deposited:

09 Jun 2015 08:27

Last Modified:

07 Nov 2015 01:30

Publisher DOI:

10.1016/j.neuropharm.2015.05.003

BORIS DOI:

10.7892/boris.69152

URI:

https://boris.unibe.ch/id/eprint/69152

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