Pyruvate as a fluorescence quencher: A new spectroscopic assay for pyruvate reactions

Toshio Ando; Hidetake Miyata

doi:10.1016/0003-2697(83)90065-9

Pyruvate as a fluorescence quencher: A new spectroscopic assay for pyruvate reactions

Toshio Ando, Hidetake Miyata

Research output: Contribution to journal › Article › peer-review

12 Citations (Scopus)

Abstract

Pyruvate ion, which is biologically ubiquitous and participates in many metabolic reactions, was found to be an effective quencher of fluorescence. Compared to other negatively charged quenchers such as I^-, pyruvate is not toxic to proteins. By adding an inert, long-lived fluorophore to systems transacting pyruvate, it is possible to estimate activity by measuring the time course of the change in pyruvate quenching of the fluorophore. The procedure is illustrated by measuring the myosin subfragment-1 ATPase activity with a high sensitivity.

Original language	English
Pages (from-to)	170-175
Number of pages	6
Journal	Analytical Biochemistry
Volume	129
Issue number	1
DOIs	https://doi.org/10.1016/0003-2697(83)90065-9
Publication status	Published - 1983 Feb 15
Externally published	Yes

Keywords

ATPase assay
charged quencher
fluorescence quenching
pyruvate
pyruvate reactions

ASJC Scopus subject areas

Biophysics
Biochemistry
Molecular Biology
Cell Biology

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10.1016/0003-2697(83)90065-9

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title = "Pyruvate as a fluorescence quencher: A new spectroscopic assay for pyruvate reactions",

abstract = "Pyruvate ion, which is biologically ubiquitous and participates in many metabolic reactions, was found to be an effective quencher of fluorescence. Compared to other negatively charged quenchers such as I-, pyruvate is not toxic to proteins. By adding an inert, long-lived fluorophore to systems transacting pyruvate, it is possible to estimate activity by measuring the time course of the change in pyruvate quenching of the fluorophore. The procedure is illustrated by measuring the myosin subfragment-1 ATPase activity with a high sensitivity.",

keywords = "ATPase assay, charged quencher, fluorescence quenching, pyruvate, pyruvate reactions",

author = "Toshio Ando and Hidetake Miyata",

year = "1983",

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doi = "10.1016/0003-2697(83)90065-9",

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pages = "170--175",

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T1 - Pyruvate as a fluorescence quencher

T2 - A new spectroscopic assay for pyruvate reactions

AU - Ando, Toshio

AU - Miyata, Hidetake

PY - 1983/2/15

Y1 - 1983/2/15

N2 - Pyruvate ion, which is biologically ubiquitous and participates in many metabolic reactions, was found to be an effective quencher of fluorescence. Compared to other negatively charged quenchers such as I-, pyruvate is not toxic to proteins. By adding an inert, long-lived fluorophore to systems transacting pyruvate, it is possible to estimate activity by measuring the time course of the change in pyruvate quenching of the fluorophore. The procedure is illustrated by measuring the myosin subfragment-1 ATPase activity with a high sensitivity.

AB - Pyruvate ion, which is biologically ubiquitous and participates in many metabolic reactions, was found to be an effective quencher of fluorescence. Compared to other negatively charged quenchers such as I-, pyruvate is not toxic to proteins. By adding an inert, long-lived fluorophore to systems transacting pyruvate, it is possible to estimate activity by measuring the time course of the change in pyruvate quenching of the fluorophore. The procedure is illustrated by measuring the myosin subfragment-1 ATPase activity with a high sensitivity.

KW - ATPase assay

KW - charged quencher

KW - fluorescence quenching

KW - pyruvate

KW - pyruvate reactions

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JO - Analytical Biochemistry

JF - Analytical Biochemistry

SN - 0003-2697

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ER -

Pyruvate as a fluorescence quencher: A new spectroscopic assay for pyruvate reactions

Abstract

Keywords

ASJC Scopus subject areas

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