The research of the Hof Fluorescence Group might be defined by following research directions:
A) Understanding Membrane Biophysics at an Atomistic Level
B) Nanoscopic segregation of lipids
C) Specific effects in lipid bilayers – ions and beyond
D) Elucidating the Dynamic/Hydration-Function Relationships in Proteins
E) Relation between structure, membrane nanoscale organisation and protein function in cells
F) Fluorescence probes and sensors
A) Understanding Membrane Biophysics at an Atomistic Level
B) Nanoscopic segregation of lipids
C) Specific effects in lipid bilayers – ions and beyond
D) Elucidating the Dynamic/Hydration-Function Relationships in Proteins
E) Relation between structure, membrane nanoscale organisation and protein function in cells
F) Fluorescence probes and sensors
A) Understanding Membrane Biophysics at an Atomistic Level
i) Protein oligomerization at the membrane.
Amyloid oligomers (Amyloid β, α-synuclein, etc.) are thought to be the entities that spark neuronal dysfunction, cell death, and Alzheimer’s or Parkinson’s disease onset. Biomembranes appear to have a distinct effect on this process, which lies in our interest. By using advanced single-molecule fluorescence techniques, we monitor the effect of particular membrane constituents (GM1, sphingomyelin, etc.) and lipid nanodomains on the oligomerization state of these peptides related to the undesired and pathological aggregate build-up (Amaro et al., Angew. Chem. 2016, Cebecauer et al., Biophys. J. 2017).
Amaro M., Šachl R., Aydogan G., Mikhalyov I.I., Vácha R., Hof M.: GM1 Ganglioside Inhibits β-Amyloid Oligomerization Induced by Sphingomyelin. Angewandte Chemie, Int. Ed., doi: 10.1002/anie.201603178.
Cebecauer M., Hof M., Amaro M.: Impact of GM1 on Membrane-mediated Aggregation/Oligomerization of β-amyloid: Unifying View. Biophys. J., doi: 10.1016/j.bpj.2017.03.009.
ii) Membrane permeabilization
We develop functional assays to monitor membrane pore formation induced by trans-membrane or membrane associated proteins (e.g. FGF2). Recently, we have developed a statistical single molecule and single vesicle assay to distinguish physiologically relevant from unspecific protein aggregation by determining the brightness of individually diffusing in-membrane oligomers and correlating the oligomer size to membrane permeability. These experiments are done on an ensemble of single vesicles in a statistical and time-dependent manner. Our findings demonstrate that quantifying oligomeric states alone does not allow for a deep understanding of the structure-function relationship of membrane-inserted oligomers (Steringer et al., elife 2017). In parallel, we investigate the role of oxidative stress in permeabilization of mitochondrial membranes by a large family of Bcl-2 proteins, which are involved in apoptosis (Dingeldein et al., Biophys. J. 2017, Lidman et al. BBA 2016).
Dingeldein A.P.G., Pokorná Š., Lidman M., Sparrman T., Šachl R., Hof M., Gröbner G.
Apoptotic Bax at Oxidatively Stressed Mitochondrial Membranes: Lipid Dynamics and Permeabilization. Biophys. J., doi: 10.1016/j.bpj.2017.04.019
Lidman M., Pokorná Š., Dingeldein A.P.G., Sparrman T., Wallgren M., Šachl R., Hof M., Gröbner G.
The oxidized phospholipid PazePC promotes permeabilization of mitochondrial membranes by Bax. Biochim. Biophys. Acta – Biomembranes, doi: 10.1016/j.bbamem.2016.03.003
Steringer J.P., Lange S., Čujová S., Dingeldein A.P.G., Pokorná Š., Lidman M., Sparrman T., Šachl R., Hof M., Gröbner G.
Apoptotic Bax at Oxidatively Stressed Mitochondrial Membranes: Lipid Dynamics and Permeabilization. Biophys. J., doi: 10.1016/j.bpj.2017.04.019
iii) Membrane fusion
Fusion of cellular membranes is a ubiquitous and vital process in living organisms. To obtain the detailed view of this complex phenomenon, we use model systems as well as plasma membranes of living cells. In eukaryotic cells, membrane fusion is mediated by so-called SNARE proteins. In Koukalová et al. Nanoscale 2018, we use a model system for membrane fusion, inspired by SNARE proteins and based on two complementary lipopeptides CPnE4 and CPnK4. Our data provide crucial insights as to how fusion is initiated and highlight the importance of both peptides in this process. In a parallel paper by Allolio et al. PNAS 2018 we investigate the entrance of arginine rich peptides into cells through induction of membrane multilamelarity and fusion. We propose a hitherto unrecognized mechanism of passive cell entry involving fusion of multilamellar structures generated by the cell-penetrating peptides.
Allolio C., Magarkar A., Jurkiewicz P., Baxová K., Javanainen M., Mason P.E., Šachl R., Cebecauer M., Hof M., Horinek D., Heinz V., Rachel R., Ziegler C.M., Schröfel A., Jungwirth P. Arginine-rich cell-penetrating peptides induce membrane multilamellarity and subsequently enter via formation of a fusion pore, PNAS, doi: 10.1073/pnas.1811520115.
Koukalová A., Pokorna Š., Boyle A.L., Mora N., Kros A., Hof M., Šachl R. Distinct roles of SNARE-mimicking lipopeptides during initial steps of membrane fusion. Nanoscale, doi: 10.1039/c8nr05730c.
i) Protein oligomerization at the membrane.
Amyloid oligomers (Amyloid β, α-synuclein, etc.) are thought to be the entities that spark neuronal dysfunction, cell death, and Alzheimer’s or Parkinson’s disease onset. Biomembranes appear to have a distinct effect on this process, which lies in our interest. By using advanced single-molecule fluorescence techniques, we monitor the effect of particular membrane constituents (GM1, sphingomyelin, etc.) and lipid nanodomains on the oligomerization state of these peptides related to the undesired and pathological aggregate build-up (Amaro et al., Angew. Chem. 2016, Cebecauer et al., Biophys. J. 2017).
Amaro M., Šachl R., Aydogan G., Mikhalyov I.I., Vácha R., Hof M.: GM1 Ganglioside Inhibits β-Amyloid Oligomerization Induced by Sphingomyelin. Angewandte Chemie, Int. Ed., doi: 10.1002/anie.201603178.
Cebecauer M., Hof M., Amaro M.: Impact of GM1 on Membrane-mediated Aggregation/Oligomerization of β-amyloid: Unifying View. Biophys. J., doi: 10.1016/j.bpj.2017.03.009.
ii) Membrane permeabilization
We develop functional assays to monitor membrane pore formation induced by trans-membrane or membrane associated proteins (e.g. FGF2). Recently, we have developed a statistical single molecule and single vesicle assay to distinguish physiologically relevant from unspecific protein aggregation by determining the brightness of individually diffusing in-membrane oligomers and correlating the oligomer size to membrane permeability. These experiments are done on an ensemble of single vesicles in a statistical and time-dependent manner. Our findings demonstrate that quantifying oligomeric states alone does not allow for a deep understanding of the structure-function relationship of membrane-inserted oligomers (Steringer et al., elife 2017). In parallel, we investigate the role of oxidative stress in permeabilization of mitochondrial membranes by a large family of Bcl-2 proteins, which are involved in apoptosis (Dingeldein et al., Biophys. J. 2017, Lidman et al. BBA 2016).
Dingeldein A.P.G., Pokorná Š., Lidman M., Sparrman T., Šachl R., Hof M., Gröbner G.
Apoptotic Bax at Oxidatively Stressed Mitochondrial Membranes: Lipid Dynamics and Permeabilization. Biophys. J., doi: 10.1016/j.bpj.2017.04.019
Lidman M., Pokorná Š., Dingeldein A.P.G., Sparrman T., Wallgren M., Šachl R., Hof M., Gröbner G.
The oxidized phospholipid PazePC promotes permeabilization of mitochondrial membranes by Bax. Biochim. Biophys. Acta – Biomembranes, doi: 10.1016/j.bbamem.2016.03.003
Steringer J.P., Lange S., Čujová S., Dingeldein A.P.G., Pokorná Š., Lidman M., Sparrman T., Šachl R., Hof M., Gröbner G.
Apoptotic Bax at Oxidatively Stressed Mitochondrial Membranes: Lipid Dynamics and Permeabilization. Biophys. J., doi: 10.1016/j.bpj.2017.04.019
iii) Membrane fusion
Fusion of cellular membranes is a ubiquitous and vital process in living organisms. To obtain the detailed view of this complex phenomenon, we use model systems as well as plasma membranes of living cells. In eukaryotic cells, membrane fusion is mediated by so-called SNARE proteins. In Koukalová et al. Nanoscale 2018, we use a model system for membrane fusion, inspired by SNARE proteins and based on two complementary lipopeptides CPnE4 and CPnK4. Our data provide crucial insights as to how fusion is initiated and highlight the importance of both peptides in this process. In a parallel paper by Allolio et al. PNAS 2018 we investigate the entrance of arginine rich peptides into cells through induction of membrane multilamelarity and fusion. We propose a hitherto unrecognized mechanism of passive cell entry involving fusion of multilamellar structures generated by the cell-penetrating peptides.
Allolio C., Magarkar A., Jurkiewicz P., Baxová K., Javanainen M., Mason P.E., Šachl R., Cebecauer M., Hof M., Horinek D., Heinz V., Rachel R., Ziegler C.M., Schröfel A., Jungwirth P. Arginine-rich cell-penetrating peptides induce membrane multilamellarity and subsequently enter via formation of a fusion pore, PNAS, doi: 10.1073/pnas.1811520115.
Koukalová A., Pokorna Š., Boyle A.L., Mora N., Kros A., Hof M., Šachl R. Distinct roles of SNARE-mimicking lipopeptides during initial steps of membrane fusion. Nanoscale, doi: 10.1039/c8nr05730c.
B) Nanoscopic segregation of lipids
Nanoscopic segregation of lipids into nanodomains have been a hot topic of the last decade as documented by our recent concise review (Cebecauer et al. Chem. Rev. 2018). The detection of these sphingomyelin- and cholesterol-enriched platforms is very demanding for their tiny dimensions and volatile character. In fact, their presence in plasma membrane is still under debate. We focus on developing new fluorescence methods that can detect and characterize these nanodomains in terms of their size, structure, concentration, interleaflet coupling and dynamics. For that purpose, we employ Förster resonance energy transfer combined with Monte Carlo simulations (MC-FRET) to estimate their dimensions and interleaflet coupling (Vinklárek et al J. Phys. Chem. Lett. 2019), antibunching assay to give a more complex portrait of lipid nanoclustering (Šachl et al BBA-Mol.Cell Res. 2015), and spot-variation-FCS (sv-FCS) and Z-scan fluorescence correlation spectroscopy (z-scan FCS) to map the lateral mobility of lipids inside and outside the nanodomains (Šachl et al., J. Phys. D – Appl. Phys. 2016).
Cebecauer M., Amaro M., Jurkiewicz P., Sarmento M., Šachl R., Cwiklik L., Hof M., Membrane Lipid Nanodomains. Chem. Rev., doi: 10.1021/acs.chemrev.8b00322.
Vinklárek I.S., Vel’as L., Riegerová P., Skála K., Mikhalyov I., Gretskaya N., Hof M., Šachl R. Experimental Evidence of the Existence of Interleaflet Coupled Nanodomains: An MC-FRET Study, J. Phys. Chem. Lett. doi: https://doi.org/10.1021/acs.jpclett.9b00390.
Šachl R., Amaro M., Aydogan G., Koukalová A., Mikhalyov I.I., Boldyrev I.A., Humpolíčková J., Hof M. On multivalent receptor activity of GM1 in cholesterol containing membranes. BBA-Molecular Cell Research, doi: 10.1016/j.bbamcr.2014.07.016.
Šachl R., Bergstrand J., Widengren J., Hof M. Fluorescence correlation spectroscopy diffusion laws in the presence of moving nanodomains. J. Phys. D-Applied Physics, doi: 10.1088/0022-3727/49/11/114002.
Nanoscopic segregation of lipids into nanodomains have been a hot topic of the last decade as documented by our recent concise review (Cebecauer et al. Chem. Rev. 2018). The detection of these sphingomyelin- and cholesterol-enriched platforms is very demanding for their tiny dimensions and volatile character. In fact, their presence in plasma membrane is still under debate. We focus on developing new fluorescence methods that can detect and characterize these nanodomains in terms of their size, structure, concentration, interleaflet coupling and dynamics. For that purpose, we employ Förster resonance energy transfer combined with Monte Carlo simulations (MC-FRET) to estimate their dimensions and interleaflet coupling (Vinklárek et al J. Phys. Chem. Lett. 2019), antibunching assay to give a more complex portrait of lipid nanoclustering (Šachl et al BBA-Mol.Cell Res. 2015), and spot-variation-FCS (sv-FCS) and Z-scan fluorescence correlation spectroscopy (z-scan FCS) to map the lateral mobility of lipids inside and outside the nanodomains (Šachl et al., J. Phys. D – Appl. Phys. 2016).
Cebecauer M., Amaro M., Jurkiewicz P., Sarmento M., Šachl R., Cwiklik L., Hof M., Membrane Lipid Nanodomains. Chem. Rev., doi: 10.1021/acs.chemrev.8b00322.
Vinklárek I.S., Vel’as L., Riegerová P., Skála K., Mikhalyov I., Gretskaya N., Hof M., Šachl R. Experimental Evidence of the Existence of Interleaflet Coupled Nanodomains: An MC-FRET Study, J. Phys. Chem. Lett. doi: https://doi.org/10.1021/acs.jpclett.9b00390.
Šachl R., Amaro M., Aydogan G., Koukalová A., Mikhalyov I.I., Boldyrev I.A., Humpolíčková J., Hof M. On multivalent receptor activity of GM1 in cholesterol containing membranes. BBA-Molecular Cell Research, doi: 10.1016/j.bbamcr.2014.07.016.
Šachl R., Bergstrand J., Widengren J., Hof M. Fluorescence correlation spectroscopy diffusion laws in the presence of moving nanodomains. J. Phys. D-Applied Physics, doi: 10.1088/0022-3727/49/11/114002.
C) Specific effects in lipid bilayers – ions and beyond
Membranes of living organisms are in permanent contact with their surroundings, including ions. To understand the importance of ions in membrane organization and function we combine fluorescence techniques with the molecular dynamics simulations (Melcrová et al., Sci. Rep. 2016, Javanainen et al., Chem. Comm. 2017). We use a similar ensemble of methods to understand at molecular level the influence of transmembrane peptides and proteins on lipid dynamics (Olšinová et al., iScience 2018).
Javanainen M., Melcrová A., Magarkar A., Jurkiewicz P., Hof M., Jungwirth P., Martinez-Seara H. Two cations, two mechanisms: interactions of sodium and calcium with zwitterionic lipid membranes. Chem. Comm. doi: 10.1039/C7CC02208E.
Melcrová A., Pokorna Š., Pullanchery S., Kohagen M., Jurkiewicz P., Hof M., Jungwirth P., Cremer P.S., Cwiklik L. The complex nature of calcium cation interactions with phospholipid bilayers. Sci. Rep. doi:10.1038/srep38035.
Olšinová M., Jurkiewicz P., Kishko I., Sýkora J., Sabó J., Hof M., Cwiklik L., Cebecauer M. Roughness of Transmembrane Helices Reduces Lipid Membrane Dynamics. iScience., doi: 10.1016/j.isci.2018.11.026.
Membranes of living organisms are in permanent contact with their surroundings, including ions. To understand the importance of ions in membrane organization and function we combine fluorescence techniques with the molecular dynamics simulations (Melcrová et al., Sci. Rep. 2016, Javanainen et al., Chem. Comm. 2017). We use a similar ensemble of methods to understand at molecular level the influence of transmembrane peptides and proteins on lipid dynamics (Olšinová et al., iScience 2018).
Javanainen M., Melcrová A., Magarkar A., Jurkiewicz P., Hof M., Jungwirth P., Martinez-Seara H. Two cations, two mechanisms: interactions of sodium and calcium with zwitterionic lipid membranes. Chem. Comm. doi: 10.1039/C7CC02208E.
Melcrová A., Pokorna Š., Pullanchery S., Kohagen M., Jurkiewicz P., Hof M., Jungwirth P., Cremer P.S., Cwiklik L. The complex nature of calcium cation interactions with phospholipid bilayers. Sci. Rep. doi:10.1038/srep38035.
Olšinová M., Jurkiewicz P., Kishko I., Sýkora J., Sabó J., Hof M., Cwiklik L., Cebecauer M. Roughness of Transmembrane Helices Reduces Lipid Membrane Dynamics. iScience., doi: 10.1016/j.isci.2018.11.026.
D) Elucidating the Dynamic/Hydration-Function Relationships in Proteins
Investigating the molecular basis of enzyme-substrate interactions that effectively contribute to the catalytic function of the enzymes is important for understanding these molecular machines. Moreover, protein engineering largely profits from this research since it brings important implications for construction of novel superior enzymes. Under this umbrella, we study two different types of proteins: ATPases (Fischermeier et al., Angew. Chem. 2017) and Haloalkane dehalogenases (Amaro et al., JACS 2015, Kokkonen et al., JACS 2018). We apply various fluorescence techniques (comprising time-dependent fluorescence shift (TDFS) or Photoinduced electron-transfer–fluorescence correlation spectroscopy (PET-FCS)) to elucidate the role of hydration and dynamics of the active site, and protein conformational dynamics in enzyme function.
Amaro M., Brezovský J., Kováčová S., Sýkora J., Bednář D., Němec V., Lišková V., Kurumbang N:P:, Beerens K., Chaloupková R., Paruch K., Hof M., Damborský J. Site-specific analysis of protein hydration based on unnatural amino Acid fluorescence. J. Am. Chem. Soc., doi: 10.1021/jacs.5b01681.
Fischermeier E., Pospíšil P., Sayed A., Hof M., Solioz M., Fahmy K. Dipolar Relaxation Dynamics at the Active Site of an ATPase Regulated by Membrane Lateral Pressure. Angew. Chem. Int. Ed., doi: 10.1002/anie.201611582.
Kokkonen P., Sykora J., Prokop Z., Ghose A., Bednar D., Amaro M., Beerens K., Bidmanova S., Slanska M., Brezovsky J., Damborsky J., Hof M. Molecular Gating of an Engineered Enzyme Captured in Real Time. J. Am. Chem. Soc., doi: 10.1021/jacs.8b09848.
Investigating the molecular basis of enzyme-substrate interactions that effectively contribute to the catalytic function of the enzymes is important for understanding these molecular machines. Moreover, protein engineering largely profits from this research since it brings important implications for construction of novel superior enzymes. Under this umbrella, we study two different types of proteins: ATPases (Fischermeier et al., Angew. Chem. 2017) and Haloalkane dehalogenases (Amaro et al., JACS 2015, Kokkonen et al., JACS 2018). We apply various fluorescence techniques (comprising time-dependent fluorescence shift (TDFS) or Photoinduced electron-transfer–fluorescence correlation spectroscopy (PET-FCS)) to elucidate the role of hydration and dynamics of the active site, and protein conformational dynamics in enzyme function.
Amaro M., Brezovský J., Kováčová S., Sýkora J., Bednář D., Němec V., Lišková V., Kurumbang N:P:, Beerens K., Chaloupková R., Paruch K., Hof M., Damborský J. Site-specific analysis of protein hydration based on unnatural amino Acid fluorescence. J. Am. Chem. Soc., doi: 10.1021/jacs.5b01681.
Fischermeier E., Pospíšil P., Sayed A., Hof M., Solioz M., Fahmy K. Dipolar Relaxation Dynamics at the Active Site of an ATPase Regulated by Membrane Lateral Pressure. Angew. Chem. Int. Ed., doi: 10.1002/anie.201611582.
Kokkonen P., Sykora J., Prokop Z., Ghose A., Bednar D., Amaro M., Beerens K., Bidmanova S., Slanska M., Brezovsky J., Damborsky J., Hof M. Molecular Gating of an Engineered Enzyme Captured in Real Time. J. Am. Chem. Soc., doi: 10.1021/jacs.8b09848.
E) Relation between structure, membrane nanoscale organization and protein function in cells
Proper communication among particular components of the immune system enables pathogen degradation. To understand this complex mechanism, we investigate regulation of CD4 and CD8 receptors in white blood cells (e.g. T-lymphocytes) (Liu et al., J. Mol. Biol. 2019, Chum et al., J. Cell. Sci. 2016). For this purpose, we apply and develop new quantitative approaches based on cutting edge super-resolution microscopy (Lukeš et al., Nat. Comm. 2017).
Chum T., Glatzová D., Kvíčalová Z., Malínský J., Brdička T., Cebecauer M. The role of palmitoylation and transmembrane domain in sorting of transmembrane adaptor proteins. Journal of Cell Science. 129, 1 (2016), 95-107.
Liu Y., Cuendet M.A., Goffin L., Šachl R., Cebecauer M., Cariolato L., Guillaume R., Reichenbach P., Irving M., Coukos G., Luescher I.F. CD8 binding of mhc-peptide complexes in CIS or trans regulates CD8+ T cell responses, J. Mol. Biol., doi:10.1016/j.jmb.2019.10.019.
Lukeš T., Glatzová D., Kvíčalová Z., Levet F., Benda A., Letschert S., Sauer M., Brdička T., Lasser T., Cebecauer M. Quantifying protein densities on cell membranes using super-resolution optical fluctuation imaging, Nature Comm., doi: 10.1038/s41467-017-01857-x.
Proper communication among particular components of the immune system enables pathogen degradation. To understand this complex mechanism, we investigate regulation of CD4 and CD8 receptors in white blood cells (e.g. T-lymphocytes) (Liu et al., J. Mol. Biol. 2019, Chum et al., J. Cell. Sci. 2016). For this purpose, we apply and develop new quantitative approaches based on cutting edge super-resolution microscopy (Lukeš et al., Nat. Comm. 2017).
Chum T., Glatzová D., Kvíčalová Z., Malínský J., Brdička T., Cebecauer M. The role of palmitoylation and transmembrane domain in sorting of transmembrane adaptor proteins. Journal of Cell Science. 129, 1 (2016), 95-107.
Liu Y., Cuendet M.A., Goffin L., Šachl R., Cebecauer M., Cariolato L., Guillaume R., Reichenbach P., Irving M., Coukos G., Luescher I.F. CD8 binding of mhc-peptide complexes in CIS or trans regulates CD8+ T cell responses, J. Mol. Biol., doi:10.1016/j.jmb.2019.10.019.
Lukeš T., Glatzová D., Kvíčalová Z., Levet F., Benda A., Letschert S., Sauer M., Brdička T., Lasser T., Cebecauer M. Quantifying protein densities on cell membranes using super-resolution optical fluctuation imaging, Nature Comm., doi: 10.1038/s41467-017-01857-x.
F) Fluorescence probes and sensors
Understanding the photophysical behavior of fluorescent probes is essential not only for the correct interpretation of the observed phenomena but also for the development of new fluorescent sensors. Therefore, we make efforts to interpret the fluorescence behavior of lipid probes at an atomistic level (e.g. revealing the sensitivity of lipophilic probes containing nitrobenzoxadiazole (NBD) to water (Filipe et al., PCCP 2019), or mapping the applicability of recent membrane probes (Amaro et al., J Phys. D.-App. Phys. 2017)). Moreover, we contribute to the development of new fluorescence dyes with superior properties, e.g. sensors for DNA-associated processes (Dziuba et al., Chem. Sci. 2016, Dziuba et al., Angew. Chem. 2016).
Amaro M., Reina F., Hof M., Eggeling Ch., Sezgin E. Laurdan and Di-4-ANEPPDHQ probe different properties of the membrane, J. Phys. D: App. Phys. doi: 10.1088/1361-6463/aa5dbc.
Dziuba D., Jurkiewicz P., Cebecauer M., Hof M., Hocek M.. Rotational Bodipy-nucleotide as an environment-sensitive fluorescence lifetime probe for DNA interactions applicable in live cell microscopy. Angew. Chem. Int. Ed., doi: 10.1002/ange.201507922R2.
Dziuba D., Pospíšil P., Matyašovský J., Brynda J., Nachtigallová D., Rulíšek L., Pohl R., Hof M., Hocek M. Solvatochromic fluorene-linked nucleoside and DNA as color-changing fluorescent probes for sensing interactions, Chem. Sci., doi: 10.1039/C6SC02548J.
Filipe H.A.L., Pokorná Š., Hof M., Amaro M., Loura L.M.S. Orientation of nitro-group governs the fluorescence lifetime of nitrobenzoxadiazole (NBD)-labeled lipids in lipid bilayers, Phys. Chem. Chem. Phys., 2019, doi: 10.1039/c8cp06064a.
Understanding the photophysical behavior of fluorescent probes is essential not only for the correct interpretation of the observed phenomena but also for the development of new fluorescent sensors. Therefore, we make efforts to interpret the fluorescence behavior of lipid probes at an atomistic level (e.g. revealing the sensitivity of lipophilic probes containing nitrobenzoxadiazole (NBD) to water (Filipe et al., PCCP 2019), or mapping the applicability of recent membrane probes (Amaro et al., J Phys. D.-App. Phys. 2017)). Moreover, we contribute to the development of new fluorescence dyes with superior properties, e.g. sensors for DNA-associated processes (Dziuba et al., Chem. Sci. 2016, Dziuba et al., Angew. Chem. 2016).
Amaro M., Reina F., Hof M., Eggeling Ch., Sezgin E. Laurdan and Di-4-ANEPPDHQ probe different properties of the membrane, J. Phys. D: App. Phys. doi: 10.1088/1361-6463/aa5dbc.
Dziuba D., Jurkiewicz P., Cebecauer M., Hof M., Hocek M.. Rotational Bodipy-nucleotide as an environment-sensitive fluorescence lifetime probe for DNA interactions applicable in live cell microscopy. Angew. Chem. Int. Ed., doi: 10.1002/ange.201507922R2.
Dziuba D., Pospíšil P., Matyašovský J., Brynda J., Nachtigallová D., Rulíšek L., Pohl R., Hof M., Hocek M. Solvatochromic fluorene-linked nucleoside and DNA as color-changing fluorescent probes for sensing interactions, Chem. Sci., doi: 10.1039/C6SC02548J.
Filipe H.A.L., Pokorná Š., Hof M., Amaro M., Loura L.M.S. Orientation of nitro-group governs the fluorescence lifetime of nitrobenzoxadiazole (NBD)-labeled lipids in lipid bilayers, Phys. Chem. Chem. Phys., 2019, doi: 10.1039/c8cp06064a.