Transmembrane electrophoresis of 8-anilino-1-naphthalenesulfonate through egg lecithin liposome membranes

Gains, Nigel; Dawson, Alan P.

doi:10.1007/bf01868625

Cited by 17 publications

(10 citation statements)

References 23 publications

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“…Acceleration of the rate of ANS-transport across bilayer membranes in the presence of valinomycin and K + has been reported by Gains and Dawson (1975). In this section, we extend this observation to 35 neutral ionophore and cation combinations and we will discuss the mechanism by which neutral ionophores catalyze ANS-transport.…”

Section: Catalysis Of Transport By 1onophoresmentioning

confidence: 67%

1-Anilino-8-naphthalenesulfonate: A fluorescent probe of ion and ionophore transport kinetics and trans-membrane asymmetry

Haynes

Simkowitz

1977

J. Membrain Biol.

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Summary. The kinetics of the transport of the 1-anilino-8-naphthalenesulfonate (ANS-, an anionic fluorescent probe of the membrane surface) across phospholipid vesicle membranes have been studied using a stopped-flow rapid kinetic technique. The method has been used to gain detailed information about the mechanism of transport of this probe and to study ionophore-mediated cation transport across the membrane. The technique has also been exploited to study differences between the inside and outside surfaces of vesicles containing pbosphatidyl choline (PC).The following is a summary of the major conclusions of this study.

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Section: Catalysis Of Transport By 1onophoresmentioning

confidence: 67%

1-Anilino-8-naphthalenesulfonate: A fluorescent probe of ion and ionophore transport kinetics and trans-membrane asymmetry

Haynes

Simkowitz

1977

J. Membrain Biol.

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“…As this reflects the change of the transmembrane electric potential difference, only the hydrolytic activity of the correctly reconstituted ATPases is measured. ANS responds very rapidly to changes in the membrane potential (ms range) [25,42,43]. Thus, the slow rise of ANS fluorescence (s range) observed here (see Fig.…”

Section: Discussionmentioning

confidence: 51%

Kinetics of ATP hydrolysis catalyzed by isolated TF1 and reconstituted TF0F1 ATPase

Rögner

Gräber

1986

Eur J Biochem

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The rate of ATP hydrolysis catalyzed by isolated TF1 and reconstituted TFoFl was measured as a function of the ATP concentration in the presence of inhibitors [ADP, Pi and 3'-O-(l-naphthoyl)ATP]. ATP hydrolysis can be described by Michaelis-Menten kinetics with K,,,(TF1) = 390 pM and K, (TFoF,) = 180 pM. The inhibition constants are for ADP Ki(TF1) = 20 pM and Ki(TFoF1) = 100 pM, for 3'-O-(l-naphthoyl)ATP Ki(TFI) = 150 pM and Ki(TFoF1) = 3 pM, and for Pi Ki(TF1) = 60 mM. From these results it is concluded that upon binding of TFo to TF1 the mechanism of ATP hydrolysis catalyzed by TFl is not changed qualitatively; however, the kinetic constants differ quantitatively.Membrane-bound ATPases of the FoFl type can catalyze proton-transport-coupled ATP synthesis/hydrolysis [ 1 -51. The hydrophilic F1 can be removed easily from the membrane and this soluble F1 has been used for studying the kinetics of ATP hydrolysis (see e. g. [6 -81). The kinetic mechanism of ATP hydrolysis, catalyzed by the soluble F1, can be regarded as a first step in the clarification of the mechanism of protontransport-coupled ATP hydrolysis catalyzed by FoFl. This implies that the binding of F1 to Fo at the membrane does not change the reaction mechanism.Using mitochondrial F1 it has been shown that under single-site conditions (i. e. ATP concentration is less than onethird of the F1 concentration) the kinetic constants for ATP binding, ATP dissociation, Pi dissociation, and ADP dissociation are similar for F1 and FoFl [9, 101. In this work the rate of ATP hydrolysis was measured as a function of the ATP concentration in the presence of the inhibitors 3'-O-(l-naphthoyl)adenosine triphosphate (N-ATP), ADP and Pi using F1 and reconstituted FoFl of the thermophilic bacterium PS3. Because of its remarkable stability [ll] this ATPase can be easily reconstituted into liposomes [12] without significant loss of enzymatic activity. The ATP analogue N-ATP has been shown earlier to be a competitive inhibitor of ATP synthesis and hydrolysis of mitochondrial [13] and chloroplast ATPase [14, 151. The results of this investigation show that the inhibition constants Correspondence to M. Rogner,

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“…They found the rate of entry to be slow, taking up to an hour to reach completion. Gains and Dawson (1975b), while confirming the slow rate of penetration of ANS into liposomes, have pointed out the danger of drawing too close an analogy between experiments performed on such widely differing systems.…”

Section: Probe Permeabilitymentioning

confidence: 91%

“…They, however, were of the opinion that the corresponding change in submitochondrial particles was almost completely caused by an enhance~t binding. Gains and Dawson (1975b) have reported that the addition of valinomycin to lecithin liposomes suspended in media containing potassium ions both accelerates the rate of penetration of ANS and leads to changes in ANS fluorescence yield even in the absence of a transmembrane potential. They suggest that these changes might be attributed to an association between ANS and a valinomycin potassium complex.…”

Section: Analysis Of Binding Plotsmentioning

confidence: 98%

An analysis of the binding of fluorescence probes in mitochondrial systems

Williams¹,

Layton²,

Johnston³

1977

J. Membrain Biol.

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Measurements of the binding of the fluorescent probes 8-anilinonaphthalene-1-sulfonate (ANS) and ethidium ions to whole and disruped mitochondria and submitochondrial particles suggest that the inner mitochondrial membrane is freely permeable to the two probes. Equations relating the binding of permeant probes to the electro-chemical balance across the membrane of vesicular systems are derived and these equations used to analyze Scatchard plots of the binding of the two probes to energized and nonenergized mitochondria and EDTA particles.

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Transmembrane electrophoresis of 8-anilino-1-naphthalenesulfonate through egg lecithin liposome membranes

Cited by 17 publications

References 23 publications

1-Anilino-8-naphthalenesulfonate: A fluorescent probe of ion and ionophore transport kinetics and trans-membrane asymmetry

1-Anilino-8-naphthalenesulfonate: A fluorescent probe of ion and ionophore transport kinetics and trans-membrane asymmetry

Kinetics of ATP hydrolysis catalyzed by isolated TF1 and reconstituted TF0F1 ATPase

An analysis of the binding of fluorescence probes in mitochondrial systems

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