Photochemical Behavior and Na+,K+ ‐ATPase Sensitivity of Voltage‐sensitive Styrylpyridinium Fluorescent Membrane Probes
Identifieur interne : 009926 ( Main/Exploration ); précédent : 009925; suivant : 009927Photochemical Behavior and Na+,K+ ‐ATPase Sensitivity of Voltage‐sensitive Styrylpyridinium Fluorescent Membrane Probes
Auteurs : Steve Amoroso [Australie] ; Vanessa V. Agen ; Thomas Starke-Peterkovic [Australie] ; Malcolm D. Mcleod [Australie] ; Hans-Jürgen Apell [Allemagne] ; Pierre Sebban [France] ; Ronald J. Clarke [Australie]Source :
- Photochemistry and Photobiology [ 0031-8655 ] ; 2006-03.
Descripteurs français
- Wicri :
- topic : Photochimie.
English descriptors
- KwdEn :
- Absorbance, Absorbance spectrum, Analogous equation, Analytical grade, Anellated hemicyanine dyes, Annine, Annine dyes, Aqueous solution, Biophys, Castle hill, Chem, Chloroform, Clarke, Conformational change, Conformational changes, Constant illumination, Continuous illumination, Cuvette holder, Differential rate equation, Dimyristoylphosphatidylcholine vesicles, Dmpc, Dmpc vesicles, Dye, Emission monochromators, Emission reactions, Equilibrium position, Excitation, Excitation spectrum, Excitation wavelength, Exciting light, Field strength, Final value, Fluorescence, Fluorescence change, Fluorescence decrease, Fluorescence emission, Fluorescence increase, Fluorescence investigations, Fluorescence lifetime, Fluorescence measurements, Fluorescence titration, Fluorescent probes, Fromherz, Glass cutoff filter, Ground state, Hormonal control, Incident light intensity, Intramolecular charge transfer, Isomerization, Isomerization reaction, Kind gift, Kinetic data, Kinetic studies, Kinetics, Light intensity, Lipid, Lipid concentration, Lipid membranes, Local intramembrane, Loew, Membrane, Membrane fragments, Membrane phase, Membrane proteins, Merck, Mercury lamp, Mercury line, Methods section, Nanosecond time scale, Neuron, Observation chamber, Optical imaging, Outer medulla, Partial reactions, Phosphorylation, Photochemical behavior, Photochemical reaction, Photochemical reactions, Photochemistry, Photoisomerization, Photon flux, Phys, Probe, Protein concentration, Pyridinium rings, Radiationless deactivation, Rate constants, Reaction mechanism, Reaction scheme, Reciprocal relaxation time, Relative fluorescence change, Relative fluorescence intensity, Relaxation time, Second time scale, Similar sensitivity, Single neurons, Solvent cage, Solvent reorganization, Steady state, Steinkopff verlag, Steve amoroso, Structure mechanism, Styryl dyes, Time function, Time scale, Trans, Trans photoisomerization, Unit time, Vesicle, Visser, Voltage sensitivity, Wavelength.
- Teeft :
- Absorbance, Absorbance spectrum, Analogous equation, Analytical grade, Anellated hemicyanine dyes, Annine, Annine dyes, Aqueous solution, Biophys, Castle hill, Chem, Chloroform, Clarke, Conformational change, Conformational changes, Constant illumination, Continuous illumination, Cuvette holder, Differential rate equation, Dimyristoylphosphatidylcholine vesicles, Dmpc, Dmpc vesicles, Dye, Emission monochromators, Emission reactions, Equilibrium position, Excitation, Excitation spectrum, Excitation wavelength, Exciting light, Field strength, Final value, Fluorescence, Fluorescence change, Fluorescence decrease, Fluorescence emission, Fluorescence increase, Fluorescence investigations, Fluorescence lifetime, Fluorescence measurements, Fluorescence titration, Fluorescent probes, Fromherz, Glass cutoff filter, Ground state, Hormonal control, Incident light intensity, Intramolecular charge transfer, Isomerization, Isomerization reaction, Kind gift, Kinetic data, Kinetic studies, Kinetics, Light intensity, Lipid, Lipid concentration, Lipid membranes, Local intramembrane, Loew, Membrane, Membrane fragments, Membrane phase, Membrane proteins, Merck, Mercury lamp, Mercury line, Methods section, Nanosecond time scale, Neuron, Observation chamber, Optical imaging, Outer medulla, Partial reactions, Phosphorylation, Photochemical behavior, Photochemical reaction, Photochemical reactions, Photochemistry, Photoisomerization, Photon flux, Phys, Probe, Protein concentration, Pyridinium rings, Radiationless deactivation, Rate constants, Reaction mechanism, Reaction scheme, Reciprocal relaxation time, Relative fluorescence change, Relative fluorescence intensity, Relaxation time, Second time scale, Similar sensitivity, Single neurons, Solvent cage, Solvent reorganization, Steady state, Steinkopff verlag, Steve amoroso, Structure mechanism, Styryl dyes, Time function, Time scale, Trans, Trans photoisomerization, Unit time, Vesicle, Visser, Voltage sensitivity, Wavelength.
Abstract
RH421 is a widely used voltage‐sensitive fluorescent membrane probe. Its exposure to continuous illumination with 577 nm light from an Hg lamp leads, however, to an increase in its steady‐state fluorescence level when bound to lipid membranes. The increase occurs on the second time scale at typical light intensities and was found to be due to a single‐photon excited‐state isomerization. Modifications to the dye structure are, therefore, necessary to increase photochemical stability and allow wider application of such dyes in kinetic studies of ion‐transporting membrane proteins. The related probe ANNINE 5, which has a rigid polycyclic structure, shows no observable photochemical reaction when bound to DMPC vesicles on irradiation with 436 nm light. The voltage sensitivity of ANNINE 5 was tested with the use of Na+,K+‐ATPase membrane fragments. As long as ANNINE 5 is excited on the far red edge of its visible absorption band, it shows a similar sensitivity to RH421 in detecting charge‐translocating reactions triggered by ATP phosphorylation. Unfortunately the wavelengths necessary for ANNINE 5 excitation are in a region where the Hg lamps routinely used in stopped‐flow apparatus have no significant lines available for excitation.
Url:
DOI: 10.1562/2005-06-08-RA-569
Affiliations:
- Allemagne, Australie, France
- Bade-Wurtemberg, District de Fribourg-en-Brisgau, Nouvelle-Galles du Sud
- Constance (Allemagne), Orsay, Sydney
- Université Paris-Sud, Université de Sydney
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Le document en format XML
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Agen</name></author><author><name sortKey="Starke Eterkovic, Thomas" sort="Starke Eterkovic, Thomas" uniqKey="Starke Eterkovic T" first="Thomas" last="Starke-Peterkovic">Thomas Starke-Peterkovic</name><affiliation wicri:level="4"><country xml:lang="fr">Australie</country><wicri:regionArea>School of Chemistry, University of Sydney, Sydney</wicri:regionArea><placeName><settlement type="city">Sydney</settlement><region type="état">Nouvelle-Galles du Sud</region><settlement type="city">Sydney</settlement></placeName><orgName type="university">Université de Sydney</orgName></affiliation></author><author><name sortKey="Mcleod, Malcolm D" sort="Mcleod, Malcolm D" uniqKey="Mcleod M" first="Malcolm D." last="Mcleod">Malcolm D. Mcleod</name><affiliation wicri:level="4"><country xml:lang="fr">Australie</country><wicri:regionArea>School of Chemistry, University of Sydney, Sydney</wicri:regionArea><placeName><settlement type="city">Sydney</settlement><region type="état">Nouvelle-Galles du Sud</region><settlement type="city">Sydney</settlement></placeName><orgName type="university">Université de Sydney</orgName></affiliation></author><author><name sortKey="Apell, Hans Rgen" sort="Apell, Hans Rgen" uniqKey="Apell H" first="Hans-Jürgen" last="Apell">Hans-Jürgen Apell</name><affiliation wicri:level="3"><country xml:lang="fr">Allemagne</country><wicri:regionArea>Fakultät für Biologie, Universität Konstanz, Konstanz</wicri:regionArea><placeName><region type="land" nuts="1">Bade-Wurtemberg</region><region type="district" nuts="2">District de Fribourg-en-Brisgau</region><settlement type="city">Constance (Allemagne)</settlement></placeName></affiliation></author><author><name sortKey="Sebban, Pierre" sort="Sebban, Pierre" uniqKey="Sebban P" first="Pierre" last="Sebban">Pierre Sebban</name><affiliation wicri:level="4"><country xml:lang="fr">France</country><wicri:regionArea>Laboratoire de Chimie‐Physique, Université Paris XI, Orsay</wicri:regionArea><placeName><settlement type="city">Orsay</settlement><settlement type="city">Orsay</settlement></placeName><orgName type="university">Université Paris-Sud</orgName></affiliation></author><author><name sortKey="Clarke, Ronald J" sort="Clarke, Ronald J" uniqKey="Clarke R" first="Ronald J." last="Clarke">Ronald J. Clarke</name><affiliation wicri:level="4"><country xml:lang="fr">Australie</country><wicri:regionArea>School of Chemistry, University of Sydney, Sydney</wicri:regionArea><placeName><settlement type="city">Sydney</settlement><region type="état">Nouvelle-Galles du Sud</region><settlement type="city">Sydney</settlement></placeName><orgName type="university">Université de Sydney</orgName></affiliation><affiliation wicri:level="1"><country wicri:rule="url">Australie</country></affiliation></author></analytic><monogr></monogr><series><title level="j" type="main">Photochemistry and Photobiology</title><title level="j" type="alt">PHOTOCHEMISTRY PHOTOBIOLOGY</title><idno type="ISSN">0031-8655</idno><idno type="eISSN">1751-1097</idno><imprint><biblScope unit="vol">82</biblScope><biblScope unit="issue">2</biblScope><biblScope unit="page" from="495">495</biblScope><biblScope unit="page" to="502">502</biblScope><biblScope unit="page-count">8</biblScope><publisher>Blackwell Publishing Ltd</publisher><pubPlace>Oxford, UK</pubPlace><date type="published" when="2006-03">2006-03</date></imprint><idno type="ISSN">0031-8655</idno></series></biblStruct></sourceDesc><seriesStmt><idno type="ISSN">0031-8655</idno></seriesStmt></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Absorbance</term><term>Absorbance spectrum</term><term>Analogous equation</term><term>Analytical grade</term><term>Anellated hemicyanine dyes</term><term>Annine</term><term>Annine dyes</term><term>Aqueous solution</term><term>Biophys</term><term>Castle hill</term><term>Chem</term><term>Chloroform</term><term>Clarke</term><term>Conformational change</term><term>Conformational changes</term><term>Constant illumination</term><term>Continuous illumination</term><term>Cuvette holder</term><term>Differential rate equation</term><term>Dimyristoylphosphatidylcholine vesicles</term><term>Dmpc</term><term>Dmpc vesicles</term><term>Dye</term><term>Emission monochromators</term><term>Emission reactions</term><term>Equilibrium position</term><term>Excitation</term><term>Excitation spectrum</term><term>Excitation wavelength</term><term>Exciting light</term><term>Field strength</term><term>Final value</term><term>Fluorescence</term><term>Fluorescence change</term><term>Fluorescence decrease</term><term>Fluorescence emission</term><term>Fluorescence increase</term><term>Fluorescence investigations</term><term>Fluorescence lifetime</term><term>Fluorescence measurements</term><term>Fluorescence titration</term><term>Fluorescent probes</term><term>Fromherz</term><term>Glass cutoff filter</term><term>Ground state</term><term>Hormonal control</term><term>Incident light intensity</term><term>Intramolecular charge transfer</term><term>Isomerization</term><term>Isomerization reaction</term><term>Kind gift</term><term>Kinetic data</term><term>Kinetic studies</term><term>Kinetics</term><term>Light intensity</term><term>Lipid</term><term>Lipid concentration</term><term>Lipid membranes</term><term>Local intramembrane</term><term>Loew</term><term>Membrane</term><term>Membrane fragments</term><term>Membrane phase</term><term>Membrane proteins</term><term>Merck</term><term>Mercury lamp</term><term>Mercury line</term><term>Methods section</term><term>Nanosecond time scale</term><term>Neuron</term><term>Observation chamber</term><term>Optical imaging</term><term>Outer medulla</term><term>Partial reactions</term><term>Phosphorylation</term><term>Photochemical behavior</term><term>Photochemical reaction</term><term>Photochemical reactions</term><term>Photochemistry</term><term>Photoisomerization</term><term>Photon flux</term><term>Phys</term><term>Probe</term><term>Protein concentration</term><term>Pyridinium rings</term><term>Radiationless deactivation</term><term>Rate constants</term><term>Reaction mechanism</term><term>Reaction scheme</term><term>Reciprocal relaxation time</term><term>Relative fluorescence change</term><term>Relative fluorescence intensity</term><term>Relaxation time</term><term>Second time scale</term><term>Similar sensitivity</term><term>Single neurons</term><term>Solvent cage</term><term>Solvent reorganization</term><term>Steady state</term><term>Steinkopff verlag</term><term>Steve amoroso</term><term>Structure mechanism</term><term>Styryl dyes</term><term>Time function</term><term>Time scale</term><term>Trans</term><term>Trans photoisomerization</term><term>Unit time</term><term>Vesicle</term><term>Visser</term><term>Voltage sensitivity</term><term>Wavelength</term></keywords><keywords scheme="Teeft" xml:lang="en"><term>Absorbance</term><term>Absorbance spectrum</term><term>Analogous equation</term><term>Analytical grade</term><term>Anellated hemicyanine dyes</term><term>Annine</term><term>Annine dyes</term><term>Aqueous solution</term><term>Biophys</term><term>Castle hill</term><term>Chem</term><term>Chloroform</term><term>Clarke</term><term>Conformational change</term><term>Conformational changes</term><term>Constant illumination</term><term>Continuous illumination</term><term>Cuvette holder</term><term>Differential rate equation</term><term>Dimyristoylphosphatidylcholine vesicles</term><term>Dmpc</term><term>Dmpc vesicles</term><term>Dye</term><term>Emission monochromators</term><term>Emission reactions</term><term>Equilibrium position</term><term>Excitation</term><term>Excitation spectrum</term><term>Excitation wavelength</term><term>Exciting light</term><term>Field strength</term><term>Final value</term><term>Fluorescence</term><term>Fluorescence change</term><term>Fluorescence decrease</term><term>Fluorescence emission</term><term>Fluorescence increase</term><term>Fluorescence investigations</term><term>Fluorescence lifetime</term><term>Fluorescence measurements</term><term>Fluorescence titration</term><term>Fluorescent probes</term><term>Fromherz</term><term>Glass cutoff filter</term><term>Ground state</term><term>Hormonal control</term><term>Incident light intensity</term><term>Intramolecular charge transfer</term><term>Isomerization</term><term>Isomerization reaction</term><term>Kind gift</term><term>Kinetic data</term><term>Kinetic studies</term><term>Kinetics</term><term>Light intensity</term><term>Lipid</term><term>Lipid concentration</term><term>Lipid membranes</term><term>Local intramembrane</term><term>Loew</term><term>Membrane</term><term>Membrane fragments</term><term>Membrane phase</term><term>Membrane proteins</term><term>Merck</term><term>Mercury lamp</term><term>Mercury line</term><term>Methods section</term><term>Nanosecond time scale</term><term>Neuron</term><term>Observation chamber</term><term>Optical imaging</term><term>Outer medulla</term><term>Partial reactions</term><term>Phosphorylation</term><term>Photochemical behavior</term><term>Photochemical reaction</term><term>Photochemical reactions</term><term>Photochemistry</term><term>Photoisomerization</term><term>Photon flux</term><term>Phys</term><term>Probe</term><term>Protein concentration</term><term>Pyridinium rings</term><term>Radiationless deactivation</term><term>Rate constants</term><term>Reaction mechanism</term><term>Reaction scheme</term><term>Reciprocal relaxation time</term><term>Relative fluorescence change</term><term>Relative fluorescence intensity</term><term>Relaxation time</term><term>Second time scale</term><term>Similar sensitivity</term><term>Single neurons</term><term>Solvent cage</term><term>Solvent reorganization</term><term>Steady state</term><term>Steinkopff verlag</term><term>Steve amoroso</term><term>Structure mechanism</term><term>Styryl dyes</term><term>Time function</term><term>Time scale</term><term>Trans</term><term>Trans photoisomerization</term><term>Unit time</term><term>Vesicle</term><term>Visser</term><term>Voltage sensitivity</term><term>Wavelength</term></keywords><keywords scheme="Wicri" type="topic" xml:lang="fr"><term>Photochimie</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">RH421 is a widely used voltage‐sensitive fluorescent membrane probe. Its exposure to continuous illumination with 577 nm light from an Hg lamp leads, however, to an increase in its steady‐state fluorescence level when bound to lipid membranes. The increase occurs on the second time scale at typical light intensities and was found to be due to a single‐photon excited‐state isomerization. Modifications to the dye structure are, therefore, necessary to increase photochemical stability and allow wider application of such dyes in kinetic studies of ion‐transporting membrane proteins. The related probe ANNINE 5, which has a rigid polycyclic structure, shows no observable photochemical reaction when bound to DMPC vesicles on irradiation with 436 nm light. The voltage sensitivity of ANNINE 5 was tested with the use of Na+,K+‐ATPase membrane fragments. As long as ANNINE 5 is excited on the far red edge of its visible absorption band, it shows a similar sensitivity to RH421 in detecting charge‐translocating reactions triggered by ATP phosphorylation. Unfortunately the wavelengths necessary for ANNINE 5 excitation are in a region where the Hg lamps routinely used in stopped‐flow apparatus have no significant lines available for excitation.</div></front></TEI><affiliations><list><country><li>Allemagne</li><li>Australie</li><li>France</li></country><region><li>Bade-Wurtemberg</li><li>District de Fribourg-en-Brisgau</li><li>Nouvelle-Galles du Sud</li></region><settlement><li>Constance (Allemagne)</li><li>Orsay</li><li>Sydney</li></settlement><orgName><li>Université Paris-Sud</li><li>Université de Sydney</li></orgName></list><tree><noCountry><name sortKey="Agen, Vanessa V" sort="Agen, Vanessa V" uniqKey="Agen V" first="Vanessa V." last="Agen">Vanessa V. Agen</name></noCountry><country name="Australie"><region name="Nouvelle-Galles du Sud"><name sortKey="Amoroso, Steve" sort="Amoroso, Steve" uniqKey="Amoroso S" first="Steve" last="Amoroso">Steve Amoroso</name></region><name sortKey="Clarke, Ronald J" sort="Clarke, Ronald J" uniqKey="Clarke R" first="Ronald J." last="Clarke">Ronald J. Clarke</name><name sortKey="Clarke, Ronald J" sort="Clarke, Ronald J" uniqKey="Clarke R" first="Ronald J." last="Clarke">Ronald J. Clarke</name><name sortKey="Mcleod, Malcolm D" sort="Mcleod, Malcolm D" uniqKey="Mcleod M" first="Malcolm D." last="Mcleod">Malcolm D. Mcleod</name><name sortKey="Starke Eterkovic, Thomas" sort="Starke Eterkovic, Thomas" uniqKey="Starke Eterkovic T" first="Thomas" last="Starke-Peterkovic">Thomas Starke-Peterkovic</name></country><country name="Allemagne"><region name="Bade-Wurtemberg"><name sortKey="Apell, Hans Rgen" sort="Apell, Hans Rgen" uniqKey="Apell H" first="Hans-Jürgen" last="Apell">Hans-Jürgen Apell</name></region></country><country name="France"><noRegion><name sortKey="Sebban, Pierre" sort="Sebban, Pierre" uniqKey="Sebban P" first="Pierre" last="Sebban">Pierre Sebban</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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