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  FCS Analysis of Protein Mobility on Lipid Monolayers

Khmelinskaia, A., Mücksch, J., Conci, F., Chwastek, G., & Schwille, P. (2018). FCS Analysis of Protein Mobility on Lipid Monolayers. Biophysical Journal, 114(10), 2444-2454. doi:10.1016/j.bpj.2018.02.031.

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 Creators:
Khmelinskaia, Alena1, Author           
Mücksch, Jonas1, Author           
Conci, Franco1, Author           
Chwastek, Grzegorz1, Author           
Schwille, Petra1, Author           
Affiliations:
1Schwille, Petra / Cellular and Molecular Biophysics, Max Planck Institute of Biochemistry, Max Planck Society, ou_1565169              

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Free keywords: FLUORESCENCE CORRELATION SPECTROSCOPY; AIR-WATER-INTERFACE; TRANSLATIONAL DIFFUSION; PHOSPHOLIPID MONOLAYERS; LATERAL DIFFUSION; CHOLERA-TOXIN; AIR/WATER INTERFACE; MEMBRANE CURVATURE; GANGLIOSIDE GM1; BILAYERSBiophysics;
 Abstract: In vitro membrane model systems are used to dissect complex biological phenomena under controlled unadulterated conditions. In this context, lipid monolayers are a powerful tool to particularly study the influence of lipid packing on the behavior of membrane proteins. Here, monolayers deposited in miniaturized fixed area-chambers, which require only minute amounts of protein, were used and shown to faithfully reproduce the characteristics of Langmuir monolayers. This assay is ideally suited to be combined with single-molecule sensitive fluorescence correlation spectroscopy (FCS) to characterize diffusion dynamics. Our results confirm the influence of lipid packing on lipid mobility and validate the use of FCS as an alternative to conventional surface pressure measurements for characterizing the monolayer. Furthermore, we demonstrate the effect of lipid density on the diffusional behavior of membrane-bound components. We exploit the sensitivity of FCS to characterize protein interactions with the lipid monolayer in a regime in which the monolayer physical properties are not altered. To demonstrate the potential of our approach, we analyzed the diffusion behavior of objects of different nature, ranging from a small peptide to a large DNA-based nanostructure. Moreover, in this work we quantify the surface viscosity of lipid monolayers. We present a detailed strategy for the conduction of point FCS experiments on lipid monolayers, which is the first step toward extensive studies of protein-monolayer interactions.

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Language(s): eng - English
 Dates: 2018
 Publication Status: Published in print
 Pages: 11
 Publishing info: -
 Table of Contents: The authors thank the Biochemistry Core Facility of the Max Planck Institute of Biochemistry for the synthesis of Alexa488 labeled MPER and for purification of the proteins MinD and MinE.
 Rev. Type: -
 Identifiers: ISI: 000432700300021
DOI: 10.1016/j.bpj.2018.02.031
 Degree: -

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Title: Biophysical Journal
  Other : Biophys. J.
Source Genre: Journal
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Publ. Info: Cambridge, Mass. : Cell Press
Pages: - Volume / Issue: 114 (10) Sequence Number: - Start / End Page: 2444 - 2454 Identifier: ISSN: 0006-3495
CoNE: https://pure.mpg.de/cone/journals/resource/954925385117