Dissecting the mechanisms of environment sensitivity of smart probes for quantitative assessment of membrane properties

Author:

Ragaller Franziska1ORCID,Andronico Luca1,Sykora Jan2,Kulig Waldemar3ORCID,Rog Tomasz3,Urem Yagmur Balim1,Abhinav 2,Danylchuk Dmytro I.4,Hof Martin2,Klymchenko Andrey4,Amaro Mariana2,Vattulainen Ilpo3ORCID,Sezgin Erdinc1ORCID

Affiliation:

1. Science for Life Laboratory, Department of Women's and Children's Health, Karolinska Institutet, 17165 Solna, Sweden

2. J. Heyrovský Institute of Physical Chemistry, Czech Academy of Sciences, Dolejškova 3, 18223 Prague 8, Czech Republic

3. Department of Physics, University of Helsinki, P.O. Box 64, FI-00014 Helsinki, Finland

4. Laboratoire de Bioimagerie et Pathologies, UMR 7021 CNRS, Université de Strasbourg, 74 Route du Rhin, 67401 Illkirch, France

Abstract

The plasma membrane, as a highly complex cell organelle, serves as a crucial platform for a multitude of cellular processes. Its collective biophysical properties are largely determined by the structural diversity of the different lipid species it accommodates. Therefore, a detailed investigation of biophysical properties of the plasma membrane is of utmost importance for a comprehensive understanding of biological processes occurring therein. During the past two decades, several environment-sensitive probes have been developed and become popular tools to investigate membrane properties. Although these probes are assumed to report on membrane order in similar ways, their individual mechanisms remain to be elucidated. In this study, using model membrane systems, we characterized the probes Pro12A, NR12S and NR12A in depth and examined their sensitivity to parameters with potential biological implications, such as the degree of lipid saturation, double bond position and configuration ( cis versus trans ), phospholipid headgroup and cholesterol content. Applying spectral imaging together with atomistic molecular dynamics simulations and time-dependent fluorescent shift analyses, we unravelled individual sensitivities of these probes to different biophysical properties, their distinct localizations and specific relaxation processes in membranes. Overall, Pro12A, NR12S and NR12A serve together as a toolbox with a wide range of applications allowing to select the most appropriate probe for each specific research question.

Funder

Syöpäsäätiö

Vetenskapsrådet

Knut och Alice Wallenbergs Stiftelse

Science for Life Laboratory

Helsinki Institute for Life Science

Sigrid Juselius Foundation

Suomen Akatemia

Ministère de la Recherche

Karolinska Institutet

Human Frontier Science Program

Publisher

The Royal Society

Subject

General Biochemistry, Genetics and Molecular Biology,Immunology,General Neuroscience

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