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Plasmalemma-associated malate dehydrogenase activity in onion root cells

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Summary

Plasma membrane vesicles isolated from onion roots showed oxaloacetate reductase activity as well as other oxidoreductase activities. Purification and further sequencing showed that the protein responsible for the activity is a 40 kDa protein which corresponds to the cytosolic soluble malate dehydrogenase. However, the activity remained bound to the membrane after repeated freezing and thawing cycles and further washing, excluding a cytosolic contamination as the source of the activity. Furthermore, a second 28 kDa protein has been copurified together with the 40 kDa protein. The plasmalemma oxaloacetate reductase activity shows both donor and acceptor sites located towards the cytoplasmic side of the plasma membrane. This enzyme catalyzed the oxidation of NADH by oxaloacetate and the reduction of NAD+ by malate in the presence of an oxaloacetate-withdrawing system. We conclude that a significant amount of the cytosolic malate dehydrogenase can be specifically attached to the cytosolic face of the plasmalemma. A possible role in a putative malate shuttle associated to the plasma membrane is discussed.

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Abbreviations

AFR:

ascorbate free radical

DQ:

duroquinone

OA:

oxaloacetate

DPIP:

dichlorophenolindophenol

MDH:

malate dehydrogenase

PHMB:

p-hydroxymercuribenzoate

References

  • Asard H, Cauberg R, Renders D, De Greef JA (1987) Duroquinone-stimulated NADH oxidase and b type cytochromes in the plasma membrane of cauliflower and mung beans. Plant Sci 53: 109–119

    Google Scholar 

  • Banaszak LJ, Bradshaw RA (1975) Malate dehydrogenases. In: Boyer PD (ed) The enzymes, vol 11, oxidation-reduction. Academic Press, New York, pp 369–396

    Google Scholar 

  • Bérczi A, Asard H (1995) NAD(P)H-utilizing oxidoreductases of the plasma membrane: an overview of presently purified proteins. Protoplasma 184: 140–144

    Google Scholar 

  • Beevers H (1979) Microbodies in higher plants. Annu Rev Plant Physiol 30: 159–193

    Google Scholar 

  • Bordier C (1981) Phase separation of integral membrane proteins in Triton X-114 solution. J Biol Chem 256: 1604–1607

    Google Scholar 

  • Borracino GS, Dipierro S, Arrigoni O (1986) Purification and properties of ascorbate free-radical reductase from potato tubers. Planta 167: 521–526

    Google Scholar 

  • Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72: 248–254

    Google Scholar 

  • Chanson A (1990) Use of pyrophosphatase activity as a reliable tonoplast marker in maize roots. Plant Sci 71: 199–207

    Google Scholar 

  • Comte J, Gautheron DC (1978) The markers of pig heart mitochondrial subfractions II: on the association of malate dehydrogenase with inner membrane. Biochimie 60: 1298–1305

    Google Scholar 

  • Cördoba-Pedregosa MC, Serrano A, Córdoba F, González-Reyes JA, Navas P, Villalba JM (1995) Topography of the 27 and 31 kDa electron transport proteins in the onion root plasma membrane. Biochim Biophys Res Commun 216: 1054–1059

    Google Scholar 

  • Delhaize E, Ryan PR (1995) Aluminum toxicity and tolerance in plants. Plant Physiol 107: 315–321

    Google Scholar 

  • Döring O, Lüthje S (1996) Molecular components and biochemistry of electron transport in plant plasma membrane. Mol Membr Biol 13: 127–142

    Google Scholar 

  • Gietl C (1992) Partitioning of malate dehydrogenase isoenzymes into glyoxysomes, mitochondria, and chloroplasts. Plant Physiol 100: 557–559

    Google Scholar 

  • Goonerwardena H, Wilson SB (1979) The oxidation of malate by isolated turnip (Brassica napus L.) mitochondria II: the malateoxidizing enzymes, number and location. J Exp Bot 30: 889–903

    Google Scholar 

  • Gross GG (1977) Cell wall-bound malate dehydrogenase from horseradish. Phytochemistry 16: 319–321

    Google Scholar 

  • —, Janse C, Elstner EF (1977) Involvement of malate, monophenols, and the superoxide radical in hydrogen peroxide formation by isolated cell walls from horseradish (Armoracia lapathifolia Gilib.). Planta 136: 271–276

    Google Scholar 

  • Hayes MK, Luethy MH, Elthon TE (1991) Mitochondrial malate dehydrogenases from corn. Plant Physiol 97: 1381–1387

    Google Scholar 

  • Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 277: 680–685

    Google Scholar 

  • Lance C, Rustin P (1984) The central role of malate in plant metabolism. Physiol Veg 22: 625–641

    Google Scholar 

  • Lord JM, Kagawa T, Moore TS, Beevers H (1973) Endoplasmic reticulum as the site of lecithin formation in castor bean endosperm. J Cell Biol 57: 659–667

    Google Scholar 

  • Luster DG, Buckhout TJ (1989) Purification and identification of a plasma membrane associated electron transport protein from maize (Zea mays L.) roots. Plant Physiol 91: 1014–1019

    Google Scholar 

  • Martinoia E, Rentsch D (1994) Malate compartmentation: responses to a complex metabolism. Annu Rev Plant Physiol Mol Biol 45: 447–467

    Google Scholar 

  • Merrill CR, Goldman D, Van Keuren ML (1984) Gel protein stains: silver stain. Methods Enzymol 104: 441–447

    Google Scholar 

  • Møller IM, Crane FL (1990) Redox processes in the plasma membrane. In: Larsson C, Møller IM (eds) The plant plasma membrane: structure, function and molecular biology. Springer, Berlin Heidelberg New York Tokyo, pp 93–126

    Google Scholar 

  • —, Fredlund KM, Bérczi A (1995) The stereospecificity, purification, and characterization of an NADH-ferricyanide reductase from spinach leaf plasma membrane. Protoplasma 184: 124–132

    Google Scholar 

  • Moore GE, Gadol SM, Robinson JB, Srere PA (1984) Binding of citrate synthase and malate dehydrogenase to mitochondrial inner membranes: tissue distribution and metabolite effects. Biochem Biophys Res Commun 121: 612–618

    Google Scholar 

  • Morré DJ, Brightman AO, Crane FL (1991) Oxidoreductase activities of the plant plasma membrane and the control of plant growth. In: Penel C, Greppin H (eds) Plant signalling, plasma membrane and change of state. Université de Genéve, Geneva, Switzerland, pp 59–77

    Google Scholar 

  • Palmgren MG, Askerlund P, Fredrikson K, Widell S, Sommarin M, Larsson C (1990a) Sealed inside-out and right-side-out plasma membrane vesicles: optimal conditions for formation and separation. Plant Physiol 92: 871–880

    Google Scholar 

  • —, Sommarin M, Ulvskov P, Larsson C (1990b) Effect of detergents on the H+-ATPase activity of inside-out and right-side-out plant plasma membrane vesicles. Biochim Biophys Acta 1221: 133–140

    Google Scholar 

  • Rubinstein B, Luster DG (1993) Plasma membrane redox activity: components and role in plant processes. Annu Rev Plant Physiol Plant Mol Biol 44: 131–155

    Google Scholar 

  • Serrano A, Villalba JM, González-Reyes JA, Navas P, Córdoba F (1994a) Two distinct NAD(P)H-dependent redox enzymes isolated from onion root plasma membranes. Biochem Mol Biol Int 32: 841–849

    Google Scholar 

  • —, Córdoba F, González-Reyes JA, Navas P, Villalba JM (1994b) Purification and characterization of two distinct NAD(P)H-dehydrogenases from onion (Allium cepa L.) root plasma membrane. Plant Physiol 106: 87–96

    Google Scholar 

  • — — —, Santos C, Navas P, Villalba JM (1995) NADH-specific dehydrogenase from onion root plasma membrane: purification and characterization. Protoplasma 184: 133–139

    Google Scholar 

  • Serrano R (1988) H+-ATPase from plasma membranes ofSaccharomyces cerevisiae andAvena sativa roots: purification and reconstitution. Methods Enzymol 157: 533–544

    Google Scholar 

  • — (1989) Structure and function of plasma membrane ATPase. Annu Rev Plant Physiol Plant Mol Biol 40: 61–94

    Google Scholar 

  • Smith AF (1983) Malate dehydrogenase. In: Bergmeyer HU, Grassl M (eds) Methods of enzymatic analysis, vol 3, oxidoreductases, transferases. Verlag Chemie, Weinheim, pp 163–176

    Google Scholar 

  • Storrie B, Madden EA (1990) Isolation of subcellular organelles. Methods Enzymol 182: 203–235

    Google Scholar 

  • Stoscheck CM (1990) Quantitation of protein. Methods Enzymol 182: 50–68

    Google Scholar 

  • Trelease RN (1984) Biogenesis of glyoxysomes. Annu Rev Plant Physiol 35: 321–347

    Google Scholar 

  • Van Gestelen P, Asard H, Cauberg RJ (1997) Solubilization and separation of a plant plasma membrane NADPH-O2 synthase from other NAD(P)H oxidoreductases. Plant Physiol 115: 543–550

    Google Scholar 

  • Widell S, Larsson C (1990) A critical evaluation of markers used in plasma membrane purification. In: Larsson C, Møller IM (eds) The plant plasma membrane. Springer, Berlin Heidelberg New York Tokyo, pp 16–43

    Google Scholar 

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Author notes

  1. F. Córdoba

    Present address: Departamento de Biología Celular, Facultad de Ciencias, Universidad de Córdoba, Avda. San Alberto Magno s/n, E-14004, Córdoba, Spain

Authors and Affiliations

  1. Departamento de Biología Celular, Facultad de Ciencias, Universidad de Córdoba, Avda. San Alberto Magno s/n, E-14004, Córdoba, Spain

    M. C. Córdoba-Pedregosa, J. A. González-Reyes, A. Serrano & J. M. Villalba

  2. Departamento de Economía y Empresa, Universidad “Pablo de Olavide”, Sevilla

    P. Navas

  3. Departamento de Ciencias Agroforestales, Universidad de Huelva, Huelva

    F. Córdoba

Authors
  1. M. C. Córdoba-Pedregosa
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  2. J. A. González-Reyes
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  3. A. Serrano
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  4. J. M. Villalba
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  5. P. Navas
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  6. F. Córdoba
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Córdoba-Pedregosa, M.C., González-Reyes, J.A., Serrano, A. et al. Plasmalemma-associated malate dehydrogenase activity in onion root cells. Protoplasma 205, 29–36 (1998). https://doi.org/10.1007/BF01279290

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  • DOI: https://doi.org/10.1007/BF01279290

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