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      Mitochondrial ATPase activity and membrane fluidity changes in rat liver in response to intoxication with Buckthorn ( Karwinskia humboldtiana)

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          Abstract

          Background

          Karwinskia humboldtiana (Kh) is a poisonous plant of the rhamnacea family. To elucidate some of the subcellular effects of Kh toxicity, membrane fluidity and ATPase activities as hydrolytic and as proton-pumping activity were assessed in rat liver submitochondrial particles. Rats were randomly assigned into control non-treated group and groups that received 1, 1.5 and 2 g/Kg body weight of dry powder of Kh fruit, respectively. Rats were euthanized at day 1 and 7 after treatment.

          Results

          Rats under Kh treatment at all dose levels tested, does not developed any neurologic symptoms. However, we detected alterations in membrane fluidity and ATPase activity. Lower dose of Kh on day 1 after treatment induced higher mitochondrial membrane fluidity than control group. This change was strongly correlated with increased ATPase activity and pH gradient driven by ATP hydrolysis. On the other hand, membrane fluidity was hardly affected on day 7 after treatment with Kh. Surprisingly, the pH gradient driven by ATPase activity was significantly higher than controls despite an diminution of the hydrolytic activity of ATPase.

          Conclusions

          The changes in ATPase activity and pH gradient driven by ATPase activity suggest an adaptive condition whereby the fluidity of the membrane is altered.

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          Most cited references79

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          The ATP synthase--a splendid molecular machine.

          P Boyer (1997)
          An X-ray structure of the F1 portion of the mitochondrial ATP synthase shows asymmetry and differences in nucleotide binding of the catalytic beta subunits that support the binding change mechanism with an internal rotation of the gamma subunit. Other structural and mutational probes of the F1 and F0 portions of the ATP synthase are reviewed, together with kinetic and other evaluations of catalytic site occupancy and behavior during hydrolysis or synthesis of ATP. Subunit function as related to proton translocation and rotational catalysis is considered. Physical demonstrations of the gamma subunit rotation have been achieved. The findings have implications for other enzymatic catalyses.
            • Record: found
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            A METHOD FOR THE COLORIMETRIC DETERMINATION OF PHOSPHORUS.

            J Sumner (1944)
            A method is described for the colorimetric determination of phosphorus, using ferrous sulfate as a reducing agent. I wish to express my gratitude to the Rockefeller Foundation for financial assistance.
              • Record: found
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              Catalytic activities of mitochondrial ATP synthase in patients with mitochondrial DNA T8993G mutation in the ATPase 6 gene encoding subunit a.

              We investigated the biochemical phenotype of the mtDNA T8993G point mutation in the ATPase 6 gene, associated with neurogenic muscle weakness, ataxia, and retinitis pigmentosa (NARP), in three patients from two unrelated families. All three carried >80% mutant genome in platelets and were manifesting clinically various degrees of the NARP phenotype. Coupled submitochondrial particles prepared from platelets capable of succinate-sustained ATP synthesis were studied using very sensitive and rapid luminometric and fluorescence methods. A sharp decrease (>95%) in the succinate-sustained ATP synthesis rate of the particles was found, but both the ATP hydrolysis rate and ATP-driven proton translocation (when the protons flow from the matrix to the cytosol) were minimally affected. The T8993G mutation changes the highly conserved residue Leu(156) to Arg in the ATPase 6 subunit (subunit a). This subunit, together with subunit c, is thought to cooperatively catalyze proton translocation and rotate, one with respect to the other, during the catalytic cycle of the F(1)F(0) complex. Our results suggest that the T8993G mutation induces a structural defect in human F(1)F(0)-ATPase that causes a severe impairment of ATP synthesis. This is possibly due to a defect in either the vectorial proton transport from the cytosol to the mitochondrial matrix or the coupling of proton flow through F(0) to ATP synthesis in F(1). Whatever mechanism is involved, this leads to impaired ATP synthesis. On the other hand, ATP hydrolysis that involves proton flow from the matrix to the cytosol is essentially unaffected.

                Author and article information

                Contributors
                sagitario_garita_mar@hotmail.com
                anacristina.ramirezanguiano@gmail.com
                genarogabriel@yahoo.com
                ew820510@gmail.com
                ljglez@yahoo.com.mx
                sandra@gmail.com
                ferminpacheco@hotmail.com
                Journal
                Biol Res
                Biol. Res
                Biological Research
                BioMed Central (London )
                0716-9760
                0717-6287
                19 March 2015
                19 March 2015
                2015
                : 48
                : 1
                : 17
                Affiliations
                [ ]Departamento de Química, Centro Universitario de Ciencias Exactas e Ingenierías, Universidad de Guadalajara, Blvd. Marcelino García Barragán 1421, 44430 Guadalajara, Jalisco Mexico
                [ ]División de Neurociencias, Laboratorio de Desarrollo, Envejecimiento y Enfermedades Neurodegenerativas, Centro de Investigación Biomédica de Occidente (CIBO), Instituto Mexicano del Seguro Social (IMSS), Sierra Mojada 800, 44340 Guadalajara, Jalisco Mexico
                Article
                8
                10.1186/s40659-015-0008-9
                4376499
                4be64fcf-0043-4fa8-ac2e-38985620bee6
                © Cid-Hernández et al.; licensee BioMed Central. 2015

                This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver ( http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

                History
                : 20 May 2014
                : 27 February 2015
                Categories
                Research Article
                Custom metadata
                © The Author(s) 2015

                karwinskia humboldtiana,atpase,mitochondria,membrane fluidity,ph gradient

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