Dual-Fluorescence <scp>l</scp>-Amino Acid Reports Insertion and Orientation of Melittin Peptide in Cell Membranes

Postupalenko, Viktoriia; Zamotaiev, Oleksandr M.; Shvadchak, Volodymyr V.; Strizhak, Aleksandr V.; Pivovarenko, Vasyl G.; Klymchenko, Andrey S.; Mély, Yves

doi:10.1021/bc400325n

Cited by 36 publications

(41 citation statements)

References 58 publications

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“…Remarkably, 3HCs has become universal probes for almost any type of biomolecular interactions. 22 Being grafted to a peptide these dyes could detect the interactions with nucleic acids, 34 proteins 35 and lipid membranes 36 (Fig. 6).…”

Section: Probes Based On Intramolecular Proton Transfermentioning

confidence: 99%

Solvatochromic and Fluorogenic Dyes as Environment-Sensitive Probes: Design and Biological Applications

2017

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Fluorescent environment-sensitive probes are specially designed dyes that change their fluorescence intensity (fluorogenic dyes) or color (e.g., solvatochromic dyes) in response to change in their microenvironment polarity, viscosity, and molecular order. The studies of the past decade, including those of our group, have shown that these molecules become universal tools in fluorescence sensing and imaging. In fact, any biomolecular interaction or change in biomolecular organization results in modification of the local microenvironment, which can be directly monitored by these types of probes. In this Account, the main examples of environment-sensitive probes are summarized according to their design concepts. Solvatochromic dyes constitute a large class of environment-sensitive probes which change their color in response to polarity. Generally, they are push-pull dyes undergoing intramolecular charge transfer. Emission of their highly polarized excited state shifts to the red in more polar solvents. Excited-state intramolecular proton transfer is the second key concept to design efficient solvatochromic dyes, which respond to the microenvironment by changing relative intensity of the two emissive tautomeric forms. Due to their sensitivity to polarity and hydration, solvatochromic dyes have been successfully applied to biological membranes for studying lipid domains (rafts), apoptosis and endocytosis. As fluorescent labels, solvatochromic dyes can detect practically any type of biomolecular interactions, involving proteins, nucleic acids and biomembranes, because the binding event excludes local water molecules from the interaction site. On the other hand, fluorogenic probes usually exploit intramolecular rotation (conformation change) as a design concept, with molecular rotors being main representatives. These probes were particularly efficient for imaging viscosity and lipid order in biomembranes as well as to light up biomolecular targets, such as antibodies, aptamers and receptors. The emerging concepts to achieve fluorogenic response to the microenvironment include ground-state isomerization, aggregation-caused quenching, and aggregation-induced emission. The ground-state isomerization exploits, for instance, polarity-dependent spiro-lactone formation in silica-rhodamines. The aggregation-caused quenching uses disruption of the self-quenched dimers and nanoassemblies of dyes in less polar environments of lipid membranes and biomolecules. The aggregation-induced emission couples target recognition with formation of highly fluorescent dye aggregates. Overall, solvatochromic and fluorogenic probes enable background-free bioimaging in wash-free conditions as well as quantitative analysis when combined with advanced microscopy, such as fluorescence lifetime (FLIM) and ratiometric imaging. Further development of fluorescent environment-sensitive probes should address some remaining problems: (i) improving their optical properties, especially brightness, photostability, and far-red to near-infrared operating range; (...

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Section: Probes Based On Intramolecular Proton Transfermentioning

confidence: 99%

Solvatochromic and Fluorogenic Dyes as Environment-Sensitive Probes: Design and Biological Applications

2017

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“…Recent technological advances in the incorporation of unnatural amino acids have greatly increased the potential for expressing intrinsically fluorescent proteins that can monitored for site-specific changes in orientation [27]. Promisingly, a 3-hydroxyflavone dye, which exhibits dual color fluorescence depending on the polarity of its environment, and acridon-2-ylalanine, a visible FRET acceptor, have been incorporated into unnatural amino acids for ensemble microscopic analysis [28, 29]. …”

Section: Choice Of Probesmentioning

confidence: 99%

Single molecule optical measurements of orientation and rotations of biological macromolecules

Shroder

Lippert

Goldman

2016

Methods Appl. Fluoresc.

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The subdomains of macromolecules often undergo large orientation changes during their catalytic cycles that are essential for their activity. Tracking these rearrangements in real time opens a powerful window into the link between protein structure and functional output. Site-specific labeling of individual molecules with polarized optical probes and measuring their spatial orientation can give insight into the crucial conformational changes, dynamics, and fluctuations of macromolecules. Here we describe the range of single molecule optical technologies that can extract orientation information from these probes, we review the relevant types of probes and labeling techniques, and we highlight the advantages and disadvantages of these technologies for addressing specific inquiries.

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“…Substitution by 3-hydroxychromone derivative of two sites in the 26 amino acid sequence of melittin, a well-studied membrane-active peptide, allowed locating these sites with respect to biomembrane based on fluorescence response to local hydration and polarity (Postupalenko et al 2013).…”

Section: Responsive Analogs Of Amino Acids and Nucleic Acid Basesmentioning

confidence: 99%

Molecular-Size Fluorescence Emitters

Demchenko

2015

Introduction to Fluorescence Sensing

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A variety of fluorescent and luminescent materials in the form of molecules, their complexes and nanoparticles are available for implementation as response units into sensing and imaging technologies and we discuss their general properties that are essential for these applications. Organic dyes were the first and remain to be the most popular emitters used with these purposes. Their labeling and responsive properties are overviewed. Natural fluorescent proteins and their engineered analogues incorporate organic fluorophores that are synthesized spontaneously from the polypeptide chain inside the living cells without the need of their intrusion, as the gene products. They resulted in revolutionary advancement in cell imaging and show good prospects in sensing and reporting on cellular processes. Organic heterocyclic compounds can incorporate metal ions generating long-living luminescence. Moreover, noble metal particles of a very small size exhibit unique light-absorbing and fluorescent emission properties. In this Chapter we focus on these types of fluorophores that possess subnanometer dimensions and display single type of absorption-emission cycle. The more complex nanostructures and nanocomposites will be overviewed in Chaps. 5 and 6 that follow. Fluorophores and Their Characteristics General Properties of Fluorescent DyesIn view of tremendous diversity of structures, chemical reactivities and spectroscopic properties, one needs certain practically useful criteria for the dye selection for the purpose of sensing and imaging. They can be formulated in the form of spectroscopic parameters that have to be optimized. It is the parameter describing the ability of molecule to absorb light at a particular wavelength. The absorbance E measured at any wavelength on spectrophotometer is proportional to the dye concentration c (in mol/l) and the light path length in a sample l (in cm). In this relation, ε plays the role of coefficient of proportionality, so that E = εcl. The value of molar absorbance is obtained by purifying and drying the dye, dissolving it at low concentration and measuring E on a spectrophotometer. The broadly used term extinction includes absorbance and light scattering. Absorbance is a unique characteristic of a molecule under certain environmental conditions. In general, the bigger the fluorophore size, the greater is the probability that the photon will be absorbed.The value of ε max at the maximum of absorption band is the common characteristic of the light-absorbing power of the dye. The dye 'brightness' that determines the absolute sensitivity of fluorescence detection (Wetzl et al. 2003) is the product of molar absorbance and quantum yield, Φ. The absorbance is related to optical cross-section and for organic dyes, ε max varies from ca. 10 3 l mol −1 cm −1 for small dyes, such as coumarins to ca. 10 5 l mol −1 cm −1 for larger fluorophores such as cyanines and phthalocyanines (Lavis and Raines 2008). It should be selected as high as possible, but the limiting factor is the fluorophore size. Us...

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Dual-Fluorescence l-Amino Acid Reports Insertion and Orientation of Melittin Peptide in Cell Membranes

Cited by 36 publications

References 58 publications

Solvatochromic and Fluorogenic Dyes as Environment-Sensitive Probes: Design and Biological Applications

Solvatochromic and Fluorogenic Dyes as Environment-Sensitive Probes: Design and Biological Applications

Single molecule optical measurements of orientation and rotations of biological macromolecules

Molecular-Size Fluorescence Emitters

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