Membrane-targeted push-pull azobenzenes for the optical modulation of membrane potential

Valentina Sesti; Arianna Magni; Matteo Moschetta; Chiara Florindi; Marlene E. Pfeffer; Mattia Lorenzo DiFrancesco; Michele Guizzardi; Giulia Folpini; Luca Sala; Alessandra Gilda Ritacca; Beatrice Campanelli; Paola Moretti; Giuseppe Maria Paternò; Luca Maragliano; Matteo Tommasini; Francesco Lodola; Elisabetta Colombo; Fabio Benfenati; Chiara Bertarelli; Guglielmo Lanzani

doi:10.1038/s41377-024-01669-x

Go to Nature Website

Your Location：

Home >

Browse articles >

Membrane-targeted push-pull azobenzenes for the optical modulation of membrane potential

Original Articles

- Membrane-targeted push-pull azobenzenes for the optical modulation of membrane potential
- Membrane-targeted push-pull azobenzenes for the optical modulation of membrane potential
- LSA 2025年14卷第1期页码：93-109
- Affiliations：
  
  1.Department of Chemistry, Materials and Chemical Engineering, "Giulio Natta" Politecnico di Milano, Milano 20133, Italy
  2.Center for Nanoscience and Technology, Istituto Italiano di Tecnologia, Milano 20134, Italy
  3.Department of Physics, Politecnico di Milano, Milano 20133, Italy
  4.Department of Biotechnology and Biosciences, University of Milano-Bicocca, Milano 20126, Italy
  5.Center for Synaptic Neuroscience and Technology, Istituto Italiano di Tecnologia, Genova 16132, Italy
  6.IRCCS Ospedale Policlinico San Martino, Genova 16132, Italy
  7.Institute for Photonics and Nanotechnologies (IFN), National Research Council (CNR), Milano 20133, Italy
  8.Istituto Auxologico Italiano IRCCS, Center for Cardiac Arrhythmias of Genetic Origin and Laboratory of Cardiovascular Genetics, Milano 20095, Italy
  9.Department of Life and Environmental Sciences, Polytechnic University of Marche, Ancona 60131, Italy
  10.Present address: Department of Materials Science and Engineering, Stanford University, Stanford, CA 94305, USA
- Author bio：
  
  Chiara Bertarelli (chiara.bertarelli@polimi.it)
  Guglielmo Lanzani (guglielmo.lanzani@iit.it)
- Funds：
- DOI：10.1038/s41377-024-01669-x
  中图分类号：
- 纸质出版日期：2025-01-31，
  
  网络出版日期：2025-01-01，
  
  收稿日期：2024-04-09，
  
  修回日期：2024-10-08，
  
  录用日期：2024-10-24
- Accepted：
Scan QR Code
Membrane-targeted push-pull azobenzenes for the optical modulation of membrane potential[J]. LSA, 2025,14(1):93-109.

Sesti, V. et al. Membrane-targeted push-pull azobenzenes for the optical modulation of membrane potential. Light: Science & Applications, 14, 93-109 (2025).
Membrane-targeted push-pull azobenzenes for the optical modulation of membrane potential[J]. LSA, 2025,14(1):93-109. DOI： 10.1038/s41377-024-01669-x.

Sesti, V. et al. Membrane-targeted push-pull azobenzenes for the optical modulation of membrane potential. Light: Science & Applications, 14, 93-109 (2025). DOI： 10.1038/s41377-024-01669-x.

摘要

Abstract

We introduce a family of membrane-targeted azobenzenes (MTs) with a push-pull character as a new tool for cell stimulation. These molecules are water soluble and spontaneously partition in the cell membrane. Upon light irradiation

they isomerize from

trans

cis

changing the local charge distribution and thus stimulating the cell response. Specifically

MTs photoisomerization induces clear and reproducible depolariz

ation. The most promising species

MTP2

was extensively studied. Time-resolved spectroscopy techniques provide insights into the excited state evolution and a complete understanding of its isomerization reaction. Molecular Dynamics simulations reveal the spontaneous and stable partitioning of the compound into the cellular membrane

without significant alterations to the bilayer thickness. MTP2 was tested in different cell types

including HEK293T cells

primary neurons

and cardiomyocytes

and a steady depolarization is always recorded. The observed membrane potential modulation in in-vitro models is attributed to the variation in membrane surface charge

resulting from the light-driven modulation of the MT dipole moment within the cell membrane. Additionally

a developed mathematical model successfully captures the temporal evolution of the membrane potential upon photostimulation. Despite being insufficient for triggering action potentials

the rapid light-induced depolarization holds potential applications

particularly in cardiac electrophysiology. Low-intensity optical stimulation with these modulators could influence cardiac electrical activity

demonstrating potential efficacy in destabilizing and terminating cardiac arrhythmias. We anticipate the MTs approach to find applications in neuroscience

biomedicine

and biophotonics

providing a tool for modulating cell physiology without genetic interventions.

关键词

Keywords

references

Fenno, L., Yizhar, O. & Deisseroth, K. The development and application of optogenetics. Annu. Rev. Neurosci. 34 , 389–412 (2011)..

Scanziani, M. & Häusser, M. Electrophysiology in the age of light. Nature 461 , 930–939 (2009)..

Thompson, A. C., Stoddart, P. R. & Jansen, E. D. Optical stimulation of neurons. Curr. Mol. Imaging 3 , 162–177 (2015)..

Di Maria, F. et al. The evolution of artificial light actuators in living systems: from planar to nanostructured interfaces. Chem. Soc. Rev. 47 , 4757–4780 (2018)..

Rau, H. Azo compounds. In Photochromism: Molecules and Systems (eds Dürr, H. & Bouas-Laurent, H. ) 165–192 (Amsterdam: Elsevier, 2003).

Beharry, A. A. & Woolley, G. A. Azobenzene photoswitches for biomolecules. Chem. Soc. Rev. 40 , 4422–4437 (2011)..

Tochitsky, I. et al. Restoring vision to the blind with chemical photoswitches. Chem. Rev. 118 , 10748–10773 (2018)..

Szymański, W. et al. Reversible photocontrol of biological systems by the incorporation of molecular photoswitches. Chem. Rev. 113 , 6114–6178 (2013)..

Bregestovski, P., Maleeva, G. & Gorostiza, P. Light-induced regulation of ligand-gated channel activity. Br. J. Pharmacol. 175 , 1892–1902 (2018)..

DiFrancesco, M. L. et al. Neuronal firing modulation by a membrane-targeted photoswitch. Nat. Nanotechnol. 15 , 296–306 (2020)..

Paternò, G. M. et al. The effect of an intramembrane light-actuator on the dynamics of phospholipids in model membranes and intact cells. Langmuir 36 , 11517–11527 (2020)..

Magni, A. et al. Azobenzene photoisomerization probes cell membrane viscosity. Phys. Chem. Chem. Phys. 24 , 8716–8723 (2022)..

Patern ò, G. M. et al. Membrane environment enables ultrafast isomerization of amphiphilic azobenzene. Adv. Sci. 7 , 1903241 (2020)..

Venturino, I. et al. Skeletal muscle cells opto-stimulation by intramembrane molecular transducers. Commun. Biol. 6 , 1148 (2023)..

Vurro, V. et al. Optical modulation of excitation-contraction coupling in human-induced pluripotent stem cell-derived cardiomyocytes. iScience 26 , 106121 (2023)..

de Souza-Guerreiro, T. C. et al. Membrane targeted azobenzene drives optical modulation of bacterial membrane potential. Adv. Sci. 10 , 2205007 (2023)..

Moschetta, M. et al. Modulation of mechanosensitive potassium channels by a membrane-targeted nongenetic photoswitch. J. Phys. Chem. B 127 , 8869–8878 (2023)..

Martinac, B., Adler, J. & Kung, C. Mechanosensitive ion channels of E . coli activated by amphipaths. Nature 348 , 261–263 (1990)..

Bassetto, C. A. Z. et al. Photolipid excitation triggers depolarizing optocapacitive currents and action potentials. Nat. Commun. 15 , 1139 (2024)..

Vurro, V. et al. Molecular design of amphiphilic plasma membrane-targeted azobenzenes for nongenetic optical stimulation. Front. Mater. 7 , 631567 (2021)..

Bandara, H. M. D. & Burdette, S. C. Photoisomerization in different classes of azobenzene. Chem. Soc. Rev. 41 , 1809–1825 (2012)..

Tulumello, D. V. & Deber, C. M. SDS micelles as a membrane-mimetic environment for transmembrane segments. Biochemistry 48 , 12096–12103 (2009)..

Lakowicz, J. R. Principles of Fluorescence Spectroscopy. 3rd edn. (New York: Springer, 2006).

Gille, K., Knoll, H. & Quitzsch, K. Rate constants of the thermal cis-trans isomerization of azobenzene dyes in solvents, acetone/water mixtures, and in microheterogeneous surfactant solutions. Int. J. Chem. Kinet. 31 , 337–350 (1999)..

Parisio, G., Ferrarini, A. & Sperotto, M. M. Model studies of lipid flip-flop in membranes. Int. J. Adv. Eng. Sci. Appl. Math. 8 , 134–146 (2016)..

Deamer, D. W. & Bramhall, J. Permeability of lipid bilayers to water and ionic solutes. Chem. Phys. Lipids 40 , 167–188 (1986)..

Wagstaff, K. M. & Jans, D. A. Protein transduction: cell penetrating peptides and their therapeutic applications. Curr. Med. Chem. 13 , 1371–1387 (2006)..

Vorobyov, I. et al. Ion-induced defect permeation of lipid membranes. Biophys. J. 106 , 586–597 (2014)..

Povilaitis, S. C. et al. Design of peptides for membrane insertion: the critical role of charge separation. J. Phys. Chem. B 126 , 6454–6463 (2022)..

Rothbard, J. B., Jessop, T. C. & Wender, P. A. Adaptive translocation: the role of hydrogen bonding and membrane potential in the uptake of guanidinium-rich transporters into cells. Adv. Drug Deliv. Rev. 57 , 495–504 (2005)..

Roux, B. The membrane potential and its representation by a constant electric field in computer simulations. Biophys. J. 95 , 4205–4216 (2008)..

Stroud, R. M. et al. Glycerol facilitator GlpF and the associated aquaporin family of channels. Curr. Opin. Struct. Biol. 13 , 424–431 (2003)..

Pinto, B. I., Bassetto, C. A. Z. & Bezanilla, F. Optocapacitance: physical basis and its application. Biophys. Rev. 14 , 569–577 (2022)..

Vurro, V. et al. Photostimulation mechanism of an amphiphilic azobenzene. IL Nuovo Cimento C 46 , 143 (2023)..

Magni, A., Vurro, V. & Lanzani, G. The effects of reiterated cell photo-stimulation with an azobenzene. IL Nuovo Cimento C 46 , 154 (2023)..

Plaksin, M. et al. Thermal transients excite neurons through universal intramembrane mechanoelectrical effects. Phys. Rev. X 8 , 011043 (2018)..

Magni, A. et al. A membrane intercalating metal-free conjugated organic photosensitizer for bacterial photodynamic inactivation. Chem. Sci. 14 , 8196–8205 (2023)..

Morstein, J., Impastato, A. C. & Trauner, D. Photoswitchable lipids. ChemBioChem 22 , 73–83 (2021)..

Hussaini, S. et al. Drift and termination of spiral waves in optogenetically modified cardiac tissue at sub-threshold illumination. eLife 10 , e59954 (2021)..

Biasci, V. et al. Optogenetic manipulation of cardiac electrical dynamics using sub-threshold illumination: dissecting the role of cardiac alternans in terminating rapid rhythms. Basic Res. Cardiol. 117 , 25 (2022)..

Marchal, G. A. et al. Optogenetic manipulation of cardiac repolarization gradients using sub-threshold illumination. Front. Physiol. 14 , 1167524 (2023)..

Manzoni, C. & Cerullo, G. Design criteria for ultrafast optical parametric amplifiers. J. Opt. 18 , 103501 (2016)..

Jo, S. et al. CHARMM-GUI: a web-based graphical user interface for CHARMM. J. Comput. Chem. 29 , 1859–1865 (2008)..

Phillips, J. C. et al. Scalable molecular dynamics with NAMD. J. Comput. Chem. 26 , 1781–1802 (2005)..

Huang, J. & Mackerell, A. D. Jr CHARMM36 all-atom additive protein force field: validation based on comparison to NMR data. J. Comput. Chem. 34 , 2135–2145 (2013)..

Vanommeslaeghe, K. et al. CHARMM general force field: a force field for drug-like molecules compatible with the CHARMM all-atom additive biological force fields. J. Comput. Chem. 31 , 671–690 (2010)..

Feller, S. E. et al. Constant pressure molecular dynamics simulation: the Langevin piston method. J. Chem. Phys. 103 , 4613–4621 (1995)..

Darden, T., York, D. & Pedersen, L. Particle mesh Ewald: an N ⋅log( N ) method for Ewald sums in large systems. J. Chem. Phys. 98 , 10089–10092 (1993)..

Guixà-González, R. et al. MEMBPLUGIN: studying membrane complexity in VMD. Bioinformatics 30 , 1478–1480 (2014)..

Humphrey, W., Dalke, A. & Schulten, K. VMD: visual molecular dynamics. J. Mol. Graph. 14 , 33–38 (1996)..

de Jong, D. H. et al. Improved parameters for the martini coarse-grained protein force field. J. Chem. Theory Comput. 9 , 687–697 (2013)..

Barnoud, J. CG builder. available on line https://jbarnoud.github.io/cgbuilder/ https://jbarnoud.github.io/cgbuilder/ .

Wassenaar, T. A. et al. Computational lipidomics with insane : a versatile tool for generating custom membranes for molecular simulations. J. Chem. Theory Comput. 11 , 2144–2155 (2015)..

Abraham, M. J. et al. GROMACS: high performance molecular simulations through multi-level parallelism from laptops to supercomputers. SoftwareX 1-2 , 19–25 (2015)..

Bussi, G., Donadio, D. & Parrinello, M. Canonical sampling through velocity rescaling. J. Chem. Phys. 126 , 014101 (2007)..

Berendsen, H. J. C. et al. Molecular dynamics with coupling to an external bath. J. Chem. Phys. 81 , 3684–3690 (1984)..

Santos, D. E. S. et al. SuAVE: a tool for analyzing curvature-dependent properties in chemical interfaces. J. Chem. Inf. Model. 60 , 473–484 (2020)..

Lolicato, M. et al. Transmembrane helix straightening and buckling underlies activation of mechanosensitive and thermosensitive K 2P channels. Neuron 84 , 1198–1212 (2014)..

Lian, X. J. et al. Directed cardiomyocyte differentiation from human pluripotent stem cells by modulating Wnt/β-catenin signaling under fully defined conditions. Nat. Protoc. 8 , 162–175 (2013)..

Sala, L. et al. Use of hiPSC-derived cardiomyocytes to rule out proarrhythmic effects of drugs: the case of hydroxychloroquine in COVID-19. Front. Physiol. 12 , 730127 (2022)..

浏览量

Downloads

CSCD

文章被引用时，请邮件提醒。

Submit

工具集

关联资源

暂无数据