Fluorescent Flippers: Small‐Molecule Probes to Image Membrane Tension in Living Systems

Chen, Xiaoxiao; Bayard, Felix; Gonzalez-Sanchis, Nerea; Pamungkas, Khurnia Krisna Puji; Sakai, Naomi; Matile, Stefan

doi:10.1002/anie.202217868

Cited by 28 publications

(101 citation statements)

References 187 publications

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“…Orthogonal to solvatochromic dyes, mechanosensitive planarizable push-pull probes (commercialized as Flipper-TR® and referred to as “Flipper” hereon) that incorporate into membranes have been developed 26–28 . These probes report on their environment by changing their fluorescence lifetime 26 .…”

Section: Introductionmentioning

confidence: 99%

Quantifying fluorescence lifetime responsiveness of environment sensitive probes for membrane fluidity measurements

Ragaller,

Sjule,

Urem

et al. 2023

Preprint

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The structural diversity of different lipid species within the membrane defines its biophysical properties such as membrane fluidity, phase transition, curvature, charge distribution and tension. Environment-sensitive probes, which change their spectral properties in response to their surrounding milieu, have greatly contributed to our understanding of such biophysical properties. To realize the full potential of these probes and to avoid misinterpretation of their spectral responses, a detailed investigation of their fluorescence characteristics in different environments is necessary. Here, we examined fluorescence lifetime of two newly developed membrane order probes, NR12S and NR12A, in response to alterations in their environments such as degree of lipid saturation, cholesterol content, double bond position and configuration and phospholipid headgroup. As comparison, we investigated lifetime sensitivity of the membrane tension probe Flipper in these environments. Applying fluorescence lifetime imaging microscopy (FLIM) in both model membranes and biological membranes, all probes distinguished membrane phases by lifetime, but exhibited different lifetime sensitivities to varying membrane biophysical properties (e.g. cholesterol). While the lifetime of Flipper is particularly sensitive to membrane cholesterol content, NR12S and NR12A lifetime is moderately sensitive to both cholesterol content and lipid acyl chains. Moreover, all probes exhibit longer lifetimes at longer emission wavelengths in membranes of any complexity. This emission-wavelength dependency results in varying lifetime resolution at different spectral regions, highly relevant for FLIM data acquisition. Our data provides valuable insights on how to perform FLIM with these probes and highlights both their potential and limitations.

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Section: Introductionmentioning

confidence: 99%

Quantifying fluorescence lifetime responsiveness of environment sensitive probes for membrane fluidity measurements

Ragaller,

Sjule,

Urem

et al. 2023

Preprint

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“…Aldehyde 7 was obtained from previously reported ether 15 by simultaneous deprotection and oxidation with DDQ (Figure a and Scheme S1). Flipper 15 was synthesized in 12 steps from commercially available starting materials following reported procedures . Weakening of the push–pull system in 15 by the aldehyde acceptor in 7 was correctly reflected by the emission changing from orange to green.…”

Section: Resultsmentioning

confidence: 99%

“…Fluorescent flippers have been introduced as small-molecule fluorescent probes to image membrane tension in living systems (Figure a–c) . For the imaging of biomembrane function, − membrane tension − is particularly interesting but also particularly demanding because physical forces are detectable only through the suprastructural changes they cause . Inspired by nature, we have considered the concept of planarizable push–pull probes to tackle this challenge .…”

Section: Introductionmentioning

confidence: 99%

“…Located at the membrane surface, variable targeting units R have to be attached to this terminus (Figure ). ,− Moreover, donors D are needed in planarized flippers II to generate a strong push–pull system accounting for large red-shifts, but they are incompatible with twisted flippers I because the decoupled donor side oxidizes . Flippers such as 1 with simple alkoxy donors are not stable for this reason (Figure d).…”

Section: Introductionmentioning

confidence: 99%

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Fluorogenic In Situ Thioacetalization: Expanding the Chemical Space of Fluorescent Probes, Including Unorthodox, Bifurcated, and Mechanosensitive Chalcogen Bonds

Chen,

Gomila,

García-Arcos

et al. 2023

JACS Au

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Progress with fluorescent flippers, small-molecule probes to image membrane tension in living systems, has been limited by the effort needed to synthesize the twisted push−pull mechanophore. Here, we move to a higher oxidation level to introduce a new design paradigm that allows the screening of flipper probes rapidly, at best in situ. Late-stage clicking of thioacetals and acetals allows simultaneous attachment of targeting units and interfacers and exploration of the critical chalcogen-bonding donor at the same time. Initial studies focus on plasma membrane targeting and develop the chemical space of acetals and thioacetals, from acyclic amino acids to cyclic 1,3-heterocycles covering dioxanes as well as dithiolanes, dithianes, and dithiepanes, derived also from classics in biology like cysteine, lipoic acid, asparagusic acid, DTT, and epidithiodiketopiperazines. From the functional point of view, the sensitivity of membrane tension imaging in living cells could be doubled, with lifetime differences in FLIM images increasing from 0.55 to 1.11 ns. From a theoretical point of view, the complexity of mechanically coupled chalcogen bonding is explored, revealing, among others, intriguing bifurcated chalcogen bonds.

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“…PFBs are excellent candidates for reacting with the thiolates of proteins on the ER surface facing the cytosol . Hence, PFBs can be modified to study the membrane surface in the ER.…”

Section: Discussionmentioning

confidence: 99%

Pentafluorobenzene: Promising Applications in Diagnostics and Therapeutics

Nair,

Seo,

Hong

et al. 2023

ACS Appl. Bio Mater.

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Pentafluorobenzene (PFB) represents a class of aromatic fluorine compounds employed exclusively across a spectrum of chemical and biological applications. PFBs are credited with developing various chemical synthesis techniques, networks and biopolymers, bioactive materials, and targeted drug delivery systems. The first part of this review delves into recent developments in PFB-derived molecules for diagnostic purposes. In the latter segment, PFB's role in the domain of theragnostic applications is discussed. The review elucidates different mechanisms and interaction strategies applied in leveraging PFBs to formulate diagnostic and theragnostic tools, substantiated by proper examples. The utilization of PFBs emerges as an enabler, facilitating manifold reactions, improving materials' properties, and even opening avenues for explorative research.

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Fluorescent Flippers: Small‐Molecule Probes to Image Membrane Tension in Living Systems

Cited by 28 publications

References 187 publications

Quantifying fluorescence lifetime responsiveness of environment sensitive probes for membrane fluidity measurements

Quantifying fluorescence lifetime responsiveness of environment sensitive probes for membrane fluidity measurements

Fluorogenic In Situ Thioacetalization: Expanding the Chemical Space of Fluorescent Probes, Including Unorthodox, Bifurcated, and Mechanosensitive Chalcogen Bonds

Pentafluorobenzene: Promising Applications in Diagnostics and Therapeutics

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