Eosin fluorescence changes during Rb+ occlusion in the Na +/K+-ATPase

We used suspensions of partially purified Na+/K +-ATPase from pig kidney to compare the effects of Rb+, as a K+ congener, on the time course and on the equilibrium values of eosin fluorescence and of Rb+ occlusion. Both sets of data were collected under identical conditions in the same enzyme prepar...

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Autores principales: Montes, M.R., González-Lebrero, R.M., Garrahan, P.J., Rossi, R.C.
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Acceso en línea:http://hdl.handle.net/20.500.12110/paper_00062960_v45_n43_p13093_Montes
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spelling todo:paper_00062960_v45_n43_p13093_Montes2023-10-03T14:04:24Z Eosin fluorescence changes during Rb+ occlusion in the Na +/K+-ATPase Montes, M.R. González-Lebrero, R.M. Garrahan, P.J. Rossi, R.C. Adenosinetriphosphate Data structures Fluorescence Positive ions Rubidium Sodium Suspensions (fluids) Congeners Eosin fluorescence Hyperbolas Occlusions Enzymes adenosine triphosphatase (potassium sodium) eosin rubidium 86 article enzyme activity enzyme kinetics enzyme purification fluorescence kidney nonhuman priority journal swine Algorithms Animals Eosine Yellowish-(YS) Fluorescence Ion Transport Kinetics Models, Chemical Na(+)-K(+)-Exchanging ATPase Protein Binding Rubidium Swine Sus scrofa We used suspensions of partially purified Na+/K +-ATPase from pig kidney to compare the effects of Rb+, as a K+ congener, on the time course and on the equilibrium values of eosin fluorescence and of Rb+ occlusion. Both sets of data were collected under identical conditions in the same enzyme preparations. The incubation media lacked ATP so that all changes led to an equilibrium distribution between enzyme conformers with and without bound eosin and with and without bound or occluded Rb+. Results showed that as Rb+ concentration was increased, the equilibrium value of fluorescence decreased and occlusion increased along rectangular hyperbolas with similar half-maximal values. The time courses of attainment of equilibrium showed an initial phase which was so quick as to fall below the time resolution of our rapid-mixing apparatus. This phase was followed by the sum of at least two exponential functions of time. In the case of fluorescence the fast exponential term accounted for a larger fraction of the time course than in the case of occlusion. Comparison between experimental and simulated results suggests that fluorescence changes express a process that is coupled to Rb+ occlusion but that is completed before occlusion reaches equilibrium. © 2006 American Chemical Society. JOUR info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by/2.5/ar http://hdl.handle.net/20.500.12110/paper_00062960_v45_n43_p13093_Montes
institution Universidad de Buenos Aires
institution_str I-28
repository_str R-134
collection Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA)
topic Adenosinetriphosphate
Data structures
Fluorescence
Positive ions
Rubidium
Sodium
Suspensions (fluids)
Congeners
Eosin fluorescence
Hyperbolas
Occlusions
Enzymes
adenosine triphosphatase (potassium sodium)
eosin
rubidium 86
article
enzyme activity
enzyme kinetics
enzyme purification
fluorescence
kidney
nonhuman
priority journal
swine
Algorithms
Animals
Eosine Yellowish-(YS)
Fluorescence
Ion Transport
Kinetics
Models, Chemical
Na(+)-K(+)-Exchanging ATPase
Protein Binding
Rubidium
Swine
Sus scrofa
spellingShingle Adenosinetriphosphate
Data structures
Fluorescence
Positive ions
Rubidium
Sodium
Suspensions (fluids)
Congeners
Eosin fluorescence
Hyperbolas
Occlusions
Enzymes
adenosine triphosphatase (potassium sodium)
eosin
rubidium 86
article
enzyme activity
enzyme kinetics
enzyme purification
fluorescence
kidney
nonhuman
priority journal
swine
Algorithms
Animals
Eosine Yellowish-(YS)
Fluorescence
Ion Transport
Kinetics
Models, Chemical
Na(+)-K(+)-Exchanging ATPase
Protein Binding
Rubidium
Swine
Sus scrofa
Montes, M.R.
González-Lebrero, R.M.
Garrahan, P.J.
Rossi, R.C.
Eosin fluorescence changes during Rb+ occlusion in the Na +/K+-ATPase
topic_facet Adenosinetriphosphate
Data structures
Fluorescence
Positive ions
Rubidium
Sodium
Suspensions (fluids)
Congeners
Eosin fluorescence
Hyperbolas
Occlusions
Enzymes
adenosine triphosphatase (potassium sodium)
eosin
rubidium 86
article
enzyme activity
enzyme kinetics
enzyme purification
fluorescence
kidney
nonhuman
priority journal
swine
Algorithms
Animals
Eosine Yellowish-(YS)
Fluorescence
Ion Transport
Kinetics
Models, Chemical
Na(+)-K(+)-Exchanging ATPase
Protein Binding
Rubidium
Swine
Sus scrofa
description We used suspensions of partially purified Na+/K +-ATPase from pig kidney to compare the effects of Rb+, as a K+ congener, on the time course and on the equilibrium values of eosin fluorescence and of Rb+ occlusion. Both sets of data were collected under identical conditions in the same enzyme preparations. The incubation media lacked ATP so that all changes led to an equilibrium distribution between enzyme conformers with and without bound eosin and with and without bound or occluded Rb+. Results showed that as Rb+ concentration was increased, the equilibrium value of fluorescence decreased and occlusion increased along rectangular hyperbolas with similar half-maximal values. The time courses of attainment of equilibrium showed an initial phase which was so quick as to fall below the time resolution of our rapid-mixing apparatus. This phase was followed by the sum of at least two exponential functions of time. In the case of fluorescence the fast exponential term accounted for a larger fraction of the time course than in the case of occlusion. Comparison between experimental and simulated results suggests that fluorescence changes express a process that is coupled to Rb+ occlusion but that is completed before occlusion reaches equilibrium. © 2006 American Chemical Society.
format JOUR
author Montes, M.R.
González-Lebrero, R.M.
Garrahan, P.J.
Rossi, R.C.
author_facet Montes, M.R.
González-Lebrero, R.M.
Garrahan, P.J.
Rossi, R.C.
author_sort Montes, M.R.
title Eosin fluorescence changes during Rb+ occlusion in the Na +/K+-ATPase
title_short Eosin fluorescence changes during Rb+ occlusion in the Na +/K+-ATPase
title_full Eosin fluorescence changes during Rb+ occlusion in the Na +/K+-ATPase
title_fullStr Eosin fluorescence changes during Rb+ occlusion in the Na +/K+-ATPase
title_full_unstemmed Eosin fluorescence changes during Rb+ occlusion in the Na +/K+-ATPase
title_sort eosin fluorescence changes during rb+ occlusion in the na +/k+-atpase
url http://hdl.handle.net/20.500.12110/paper_00062960_v45_n43_p13093_Montes
work_keys_str_mv AT montesmr eosinfluorescencechangesduringrbocclusioninthenakatpase
AT gonzalezlebrerorm eosinfluorescencechangesduringrbocclusioninthenakatpase
AT garrahanpj eosinfluorescencechangesduringrbocclusioninthenakatpase
AT rossirc eosinfluorescencechangesduringrbocclusioninthenakatpase
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