scholarly journals Single-molecule fluorescence vistas of how lipids regulate membrane proteins

2021 ◽  
Author(s):  
Alyssa E. Ward ◽  
Yujie Ye ◽  
Jennifer A. Schuster ◽  
Shushu Wei ◽  
Francisco N. Barrera
Keyword(s):  
Membrane Proteins ◽  
Physical Properties ◽  
Lipid Bilayers ◽  
Single Molecule ◽  
Short Review ◽  
Single Molecule Fluorescence ◽  
Determining Factors ◽  
Fluorescence Methods ◽  
Basic Understanding ◽  
Sample Heterogeneity

The study of membrane proteins is undergoing a golden era, and we are gaining unprecedented knowledge on how this key group of proteins works. However, we still have only a basic understanding of how the chemical composition and the physical properties of lipid bilayers control the activity of membrane proteins. Single-molecule (SM) fluorescence methods can resolve sample heterogeneity, allowing to discriminate between the different molecular populations that biological systems often adopt. This short review highlights relevant examples of how SM fluorescence methodologies can illuminate the different ways in which lipids regulate the activity of membrane proteins. These studies are not limited to lipid molecules acting as ligands, but also consider how the physical properties of the bilayer can be determining factors on how membrane proteins function.

scholarly journals HDL particles incorporate into lipid bilayers – a combined AFM and single molecule fluorescence microscopy study

2017 ◽  
Vol 7 (1) ◽  
Author(s):  
Birgit Plochberger ◽  
Clemens Röhrl ◽  
Johannes Preiner ◽  
Christian Rankl ◽  
Mario Brameshuber ◽  
...  
Keyword(s):  
Fluorescence Microscopy ◽  
Lipid Bilayers ◽  
Single Molecule ◽  
Microscopy Study ◽  
Single Molecule Fluorescence ◽  
Single Molecule Fluorescence Microscopy

scholarly journals Single molecule fluorescence for membrane proteins

Methods ◽  
2018 ◽  
Vol 147 ◽  
pp. 221-228 ◽  
Author(s):  
Oliver K. Castell ◽  
Patricia M. Dijkman ◽  
Daniel N. Wiseman ◽  
Alan D. Goddard
Keyword(s):  
Membrane Proteins ◽  
Single Molecule ◽  
Single Molecule Fluorescence

Single-Molecule Fluorescence Imaging of Peptide Binding to Supported Lipid Bilayers

2009 ◽  
Vol 81 (13) ◽  
pp. 5130-5138 ◽  
Author(s):  
Christopher B. Fox ◽  
Joshua R. Wayment ◽  
Grant A. Myers ◽  
Scott K. Endicott ◽  
Joel M. Harris
Keyword(s):  
Fluorescence Imaging ◽  
Lipid Bilayers ◽  
Single Molecule ◽  
Peptide Binding ◽  
Supported Lipid Bilayers ◽  
Single Molecule Fluorescence

scholarly journals Non-vesicular transfer of membrane proteins from nanoparticles to lipid bilayers

2011 ◽  
Vol 137 (2) ◽  
pp. 217-223 ◽  
Author(s):  
Sourabh Banerjee ◽  
Crina M. Nimigean
Keyword(s):  
Membrane Proteins ◽  
Lipid Bilayer ◽  
Lipid Bilayers ◽  
Single Molecule ◽  
Lipid Membranes ◽  
Single Channel ◽  
Black Lipid Membranes ◽  
Potassium Channel Kcsa ◽  
Lipid Environment ◽  
Biochemical Preparation

Discoidal lipoproteins are a novel class of nanoparticles for studying membrane proteins (MPs) in a soluble, native lipid environment, using assays that have not been traditionally applied to transmembrane proteins. Here, we report the successful delivery of an ion channel from these particles, called nanoscale apolipoprotein-bound bilayers (NABBs), to a distinct, continuous lipid bilayer that will allow both ensemble assays, made possible by the soluble NABB platform, and single-molecule assays, to be performed from the same biochemical preparation. We optimized the incorporation and verified the homogeneity of NABBs containing a prototypical potassium channel, KcsA. We also evaluated the transfer of KcsA from the NABBs to lipid bilayers using single-channel electrophysiology and found that the functional properties of the channel remained intact. NABBs containing KcsA were stable, homogeneous, and able to spontaneously deliver the channel to black lipid membranes without measurably affecting the electrical properties of the bilayer. Our results are the first to demonstrate the transfer of a MP from NABBs to a different lipid bilayer without involving vesicle fusion.

Surfing on a new wave of single-molecule fluorescence methods

2010 ◽  
Vol 7 (3) ◽  
pp. 031001 ◽  
Author(s):  
Johannes Hohlbein ◽  
Kristofer Gryte ◽  
Mike Heilemann ◽  
Achillefs N Kapanidis
Keyword(s):  
Single Molecule ◽  
New Wave ◽  
Single Molecule Fluorescence ◽  
Fluorescence Methods

scholarly journals Single-molecule imaging with cell-derived nanovesicles reveals early binding dynamics at a cyclic nucleotide-gated ion channel

2021 ◽  
Author(s):  
Vishal R Patel ◽  
Arturo M Salinas ◽  
Darong Qi ◽  
Shipra Gupta ◽  
David J Sidote ◽  
...  
Keyword(s):  
Membrane Proteins ◽  
Cell Membrane ◽  
Ion Channel ◽  
Single Molecule ◽  
Cyclic Nucleotide ◽  
Sensory Transduction ◽  
Single Molecule Fluorescence ◽  
Native Cell ◽  
Pore Opening ◽  
Intracellular Domains

Ligand binding to membrane proteins is critical for many biological signaling processes. However, individual binding events are rarely directly observed, and their asynchronous dynamics are occluded in ensemble-averaged measures. For membrane proteins, single-molecule approaches that resolve these dynamics are challenged by dysfunction in nonnative lipid environments, lack of access to intracellular sites, and costly sample preparation. Here, we introduce an approach combining cell-derived nanovesicles, microfluidics, and single-molecule fluorescence colocalization microscopy to track individual binding events at a cyclic nucleotide-gated TAX-4 ion channel critical for sensory transduction. Our observations reveal dynamics of both nucleotide binding and a subsequent conformational change likely preceding pore opening. We further show that binding of the second ligand in the tetrameric channel is less cooperative than previously estimated from ensemble-averaged binding measures. This approach is broadly applicable to studies of binding dynamics for proteins with extracellular or intracellular domains in native cell membrane.

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