Continuous measurement of mitochondrial pH gradients in isolated hepatocytes by difference ratio spectroscopy.


:The pH gradient, delta pH, present across the inner mitochondrial membrane in isolated rat hepatocytes was continuously monitored with a novel spectroscopic technique that utilizes the weak acid fluorescein. Unlike most cytosolic pH indicators, such as 2',7'-bis(carboxyethyl)-5,(6)-carboxyfluorescein (BCECF), fluorescein freely distributes between the cytosolic and mitochondrial compartments. As is typical for weak acids, the distribution between these two compartments is governed by the magnitude of the pH gradient. Since fluorescein has two ionizable groups, the fluorescein dianion is concentrated in the mitochondrial compartment 100-fold per delta pH unit. In this compartment, fluorescein absorbance (or excitation) spectra are red-shifted about 6-8 nm in the matrix environment, as compared to the cytosolic dye at equivalent pH values. The combination of favorable mitochondrial accumulation and red-shifted spectra enables mitochondrial pH to be continuously monitored qualitatively in whole cells by dual wavelength spectroscopy (510 minus 540 nm). When the cytosolic pH is determined by independent means, the mitochondrial pH can be quantitated, based on the theoretical dependence of the fluorescein distribution ratio on delta pH, the ratio of cytosolic to mitochondrial volumes, and the known extinction coefficients for the dye in the cytosolic and mitochondrial compartments. The sensitivity of the method for following kinetic responses in mitochondrial pH is especially noteworthy; a 0.1-unit change in delta pH is easily distinguished, with a time resolution of less than a second.


Arch Biochem Biophys


Thomas PJ,Gaspers LD,Pharr C,Thomas JA




Has Abstract


1991-07-01 00:00:00














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    journal_title:Archives of biochemistry and biophysics

    pub_type: 杂志文章


    authors: Condell RA,Tappel AL

    更新日期:1983-06-01 00:00:00

  • Regulators in the DNA damage response.

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    journal_title:Archives of biochemistry and biophysics

    pub_type: 杂志文章,评审


    authors: Liu Y,Li Y,Lu X

    更新日期:2016-03-15 00:00:00

  • Regulation of 12-lipoxygenase in rat intestinal epithelial cells during differentiation and apoptosis induced by sodium butyrate.

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    journal_title:Archives of biochemistry and biophysics

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    authors: Kamitani H,Ikawa H,Hsi LC,Watanabe T,DuBois RN,Eling TE

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  • A model for hydropathy-based peptide interactions.

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    journal_title:Archives of biochemistry and biophysics

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    authors: Markus G,Tritsch GL,Parthasarathy R

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  • Purification and characterization of alanine aminotransferase from Panicum miliaceum leaves.

    abstract::Three alanine aminotransferases, two minor (AlaAT-1, AlaAT-3) and one major (AlaAT-2), were detected by native gel electrophoresis of leaf extracts from Panicum miliaceum L. AlaAT-2 was purified to homogeneity and a specific polyclonal antibody was raised against it which did not react with the other two forms of the ...

    journal_title:Archives of biochemistry and biophysics

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    authors: Son D,Jo J,Sugiyama T

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  • Interaction between the SH2 domains of ZAP-70 and the tyrosine-based activation motif 1 sequence of the zeta subunit of the T-cell receptor.

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    authors: Labadia ME,Jakes S,Grygon CA,Greenwood DJ,Schembri-King J,Lukas SM,Warren TC,Ingraham RH

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  • L-carnitine delays the killing of cultured hepatocytes by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine.

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    journal_title:Archives of biochemistry and biophysics

    pub_type: 杂志文章


    authors: Snyder JW,Kyle ME,Ferraro TN

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  • Cloning and characterization of an S-formylglutathione hydrolase from Arabidopsis thaliana.

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    authors: Kordic S,Cummins I,Edwards R

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    authors: Tatsumi K,Yamada H,Kitamura S

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  • Physiological serum copper concentrations found in malignancies cause unfolding induced aggregation of human serum albumin in vitro.

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    authors: Masaki N,Thomas AP,Hoek JB,Farber JL

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    authors: Grant RS,Passey R,Matanovic G,Smythe G,Kapoor V

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    authors: Akerboom TP,Sies H,Ziegler DM

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    authors: Thomas CJ,Surolia A

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    authors: Hering TM,Kollar J,Huynh TD,Varelas JB,Sandell LJ

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    authors: Ishii Y,Lehrer SS,Ohnishi ST,Rubin E

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    journal_title:Archives of biochemistry and biophysics

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    authors: Wang KK,Du YS,Diglio C,Tsang W,Kuo TH

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    journal_title:Archives of biochemistry and biophysics

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    authors: Perizzolo KE,Sullivan S,Waugh DF

    更新日期:1985-03-01 00:00:00

  • Transcriptional regulation of rat liver glutathione S-transferase genes by phenobarbital and 3-methylcholanthrene.

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    journal_title:Archives of biochemistry and biophysics

    pub_type: 杂志文章


    authors: Ding VD,Pickett CB

    更新日期:1985-08-01 00:00:00

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    journal_title:Archives of biochemistry and biophysics

    pub_type: 杂志文章


    authors: Stiborová M,Borek-Dohalská L,Hodek P,Mráz J,Frei E

    更新日期:2002-07-01 00:00:00

  • NMR evidence for perturbation of the copper coordination sphere upon chemical modification of arginine 141 in bovine Cu,Zn superoxide dismutase.

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    journal_title:Archives of biochemistry and biophysics

    pub_type: 杂志文章


    authors: Paci M,Desideri A,Sette M,Rotilio G

    更新日期:1991-04-01 00:00:00

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    journal_title:Archives of biochemistry and biophysics

    pub_type: 杂志文章


    authors: Saavedra C,Araneda S,Cardemil E

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  • Synthetic ceramides induce growth arrest or apoptosis by altering cellular redox status.

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    journal_title:Archives of biochemistry and biophysics

    pub_type: 杂志文章


    authors: Phillips DC,Allen K,Griffiths HR

    更新日期:2002-11-01 00:00:00