Skip to main content
SpringerLink
Log in

Pyridine nucleotide-dependent ferricyanide reduction associated with isolated plasma membranes of maize (Zea mays L.) roots

  • Published:
Protoplasma Aims and scope Submit manuscript

Summary

Plasma membranes (PM) from maize roots (Zea mays L.) were isolated by aqueous two-phase partitioning. The isolated membrane fraction showed a 4.6-fold enrichment in specific activity of the PM marker enzyme vanadate-sensitive, Mg2+-ATPase over a microsomal pellet collected at 50,000 × g. Activities of marker enzymes for mitochondria, endoplasmic reticulum, tonoplast, and Golgi apparatus were low or not detectable in the PM fraction. Quantitative morphometric analysis using the PM-specific silicotungstic acid stain showed the fraction to be > 92% PM vesicles. Using detergent stimulation of ATPase activity as a measure of structurally linked latency, greater than 90% of the PM vesicles were oriented with the cytoplasmic surface inside.

An electron transport activity was investigated in the PM fraction. The rate of NADH oxidation in the absence of an artificial electron acceptor was < 167pkat·mg protein−1; however, NADH catalysed the reduction of a variety of artificial electron acceptors including ferricyanide (2.6 nkat·mg protein−1), cytochromec (0.8 nkat·mg protein−1), a tetrazolium derivative (0.6 nkat·mg protein−1) and dichlorophenol indophenol (0.4 nkat·mg protein−1). While the NADH-dependent ferricyanide and dichlorophenol indophenol reductases were stimulated ⩾ 6-fold by 0.025% (v/v) Triton X-100, the cytochromec and INT reductases were not greatly stimulated. Washing membranes with high salt significantly decreased the NADH-dependent, and eliminated the NADPH-dependent, ferricyanide reductase activity measured in the absence of detergent. These results suggest that NADH was oxidized on the extracytoplasmic surface of the membrane; however, a significant portion of this activity was extrinsic and may have originated from cytoplasmic contamination during isolation. The greater portion of the PM-associated NAD(P)H oxidation and/or ferricyanide reduction was catalyzed on sites not exposed to the outer surface of the membrane.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

Abbreviations

BTP:

1,3-bis[tris(hydroxymethyl)-methylamino]-propane

CHAPS:

3-[(3-cholamidopropyl)dimethylammonio]-1-propane sulfonate dihydrate

cytc :

cytochromec

DCIP:

2,6-dichlorophenol indolphenol

INT:

2-(4-iodophenyl)-3-(4-nitrophenyl)-5-phenyltetrazolium chloride

kat:

mole·s−1

Mes:

2-(N-morpholino)ethanesulfonic acid

MF:

microsomal fraction

PM:

plasma membrane

STA:

silicotungstic acid

Tris:

2-amino-2-(hydroxymethyl)-1,3-propanediol

References

  • Albertsson P-Å, Andersson B, Larsson C., Åkerlund H-E (1981) Phase partition—a method for purification and analysis of cell organelles and membranes vesicles. Meth Biochemical Analysis 28: 115–150

    Google Scholar 

  • Barr R, Safranski K, Sun IL, Crane FL, Morré DJ (1984) An electrogenic proton pump associated with the Golgi apparatus of mouse liver driven by NADH and ATP. J Biol Chem 259: 14064–14067

    PubMed  Google Scholar 

  • —,Craig TA, Crane FL (1985 a) Transmembrane ferricyanide reduction in carrot cells. Biochim Biophys Acta 812: 49–54

    PubMed  Google Scholar 

  • —,Sandelius AS, Crane FL, Morré DJ (1985 b) Oxidation of reduced pyridine nucleotides by plasma membranes of soybean hypocotyls. Biochem Biophys Res Commun 131: 943–948

    PubMed  Google Scholar 

  • Buckhout TJ, Heyder-Caspers L, Sievers A (1982) Fractionation and characterization of cellular membranes from root tips of garden cress (Lepidium sativum L.). Planta 152: 108–116

    Google Scholar 

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

    PubMed  Google Scholar 

  • Crane FL, Sun IL, Clark MG, Grebing C, Löw H (1985) Transplasma-membrane redox systems in growth and development. Biochim Biophys Acta 881: 233–264

    Google Scholar 

  • De Luca L, Bader U, Hertel R, Pupillo P (1984) Detergent activity of NADH oxidase in vesicles derived from the plasma-membrane ofCucurbita pepo L. Plant Sci Lett 36: 93–98

    Google Scholar 

  • Frederico R, Giartosio CE (1983) A transplasmamembrane electron transport system in maize roots. Plant Physiol 73: 182–184

    Google Scholar 

  • Grebing C, Crane FL, Löw H, Hall K (1984) A transmembranous NADH-dehydrogenase in human erythrocyte membranes. J Bioenerg Biomemb 16: 517–533

    Google Scholar 

  • Hodges TK, Leonard RT (1972) Purification of a plasma membrane-bound adenosine triphosphatase from plant roots. Methods Enzymol 32: 392–406

    Google Scholar 

  • Kjellbom P, Larsson C (1984) Preparation and polypeptide composition of chlorophyll-free plasma membranes from leaves of light-grown spinach and barley. Physiol Plant 62: 501–509

    Google Scholar 

  • Larsson C (1983) Partition in aqueous polymer two-phase systems: a rapid method for separation of membrane particles according to their surface properties. In:Hall JL, Moore AL (eds) Isolation of membranes and organelles from plant cells. Academic Press, London New York Paris, pp 277–309

    Google Scholar 

  • —,Kjellbom P, Widell S, Lundborg T (1984) Sideness of plant plasma membrane vesicles purified by partitioning in aqueous two phase systems. FEBS Lett 171: 217–276

    Google Scholar 

  • Lin W (1984) Further characterization on the transport property of plasmalemma NADH oxidation system in isolated corn root protoplasts. Plant Physiol 74: 219–222

    Google Scholar 

  • —, (1982) Responses of corn root protoplasts to exogenous reduced nicotinamide adenine dinucleotide: oxygen consumption, ion uptake and membrane potential. Proc Natl Acad Sci USA 79: 3773–3776

    Google Scholar 

  • Luster DG, Donaldson RP (1985) Characterization of electron transport activities of the glyoxysomal membrane. Plant Physiol 77: 147

    Google Scholar 

  • Misra PC, Craig TA, Crane FL (1984) A link between transport and plasma membrane redox system(s) in carrot cells. J Bioenerg Biomembr 16: 143–152

    PubMed  Google Scholar 

  • Møller IM, Lin W (1986) Membrane-bound NAD(P)H dehydrogenases in higher plant cells. Ann Rev Plant Physiol 37: 309–334

    Google Scholar 

  • Moreau F, Jacob J-L, Dupont J, Lance C (1975) Electron transport in the membrane of lutiods from the latex ofHevea brasiliensis. Biochim Biophys Acta 396: 116–124

    PubMed  Google Scholar 

  • Morré DJ, Vigil EL, Frantz C, Goldenberg H, Crane FL (1978) Cytochemical demonstration of glutaraldehyde-resistant NADH-ferricyanide oxido-reductase activities in rat-liver plasma membranes and Golgi apparatus. Cytobiologie 18: 213–230

    PubMed  Google Scholar 

  • Pennington RJ (1961) Biochemistry of dystrophic muscles. Mitochondrial succinate-tetrazolium reductase and adenosine triphosphatase. Biochem J 80: 649–654

    PubMed  Google Scholar 

  • Pupillo P, Valenti V, De Luca L, Hertel R (1986) Kinetic characterization of reduced pyridine nucleotide dehydrogenases (duroquinone-dependent) inCucurbita microsomes. Plant Physiol 80: 384–389

    Google Scholar 

  • Qiu Z-S, Rubinstein B, Stern AI (1985) Evidence for electron transport across the plasma membrane ofZea mays root cells. Planta 165: 383–391

    Google Scholar 

  • Quail PH (1979) Plant cell fractionation. Ann Rev Plant Physiol 30: 425–484

    Google Scholar 

  • Reynolds ES (1963) The use of lead citrate at high pH as an electronopaque stain in electron microscopy. J Cell Biol 17: 208–212

    PubMed  Google Scholar 

  • Roland J-C (1978) General preparation and staining of thin sections. In:Hall JL (ed) Electron microscopy and cytochemistry of plant cells. Elsevier, Amsterdam Oxford New York, pp 1–62

    Google Scholar 

  • Roland J-C, Lembi CA, Morré DJ (1972) Phosphotungstic acidchromic acid as a selective electron-dense stain for plasma membranes of plant cells. Stain Technology 47: 195–200

    PubMed  Google Scholar 

  • Rubinstein B, Stern AI, Stout RG (1984) Redox activity at the surface of oat root cells. Plant Physiol 76: 386–391

    Google Scholar 

  • Scherer GFE, Morré DJ (1978) In vitro stimulation by 2,4-dichlorophenoxyacetic acid of an ATPase and inhibition of phosphatidate phosphatase of plant membranes. Biochem Biophys Res Comm 84: 238–247

    PubMed  Google Scholar 

  • Segel IH (1975) Enzyme kinetics. Behavior and analysis of rapid equilibrium and steady-state enzyme systems. John Wiley & Sons, New York London Sydney Toronto

    Google Scholar 

  • Sijmons PC, Van Den Briel W, Bienfait HF (1984) Cytosolic NADPH is the electron donor for extracellular Fe3+ reduction in iron-deficient bean roots. Plant Physiol 75: 219–221

    Google Scholar 

  • Thom M, Maretzki A (1985) Evidence for a plasmalemma redox system in sugarcane. Plant Physiol 77: 873–876

    Google Scholar 

  • Weibel ER, Kistler GS, Scherle WF (1966) Practical stereological methods for morphometric cytology. J Cell Biol 30: 23–38

    PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Plant Photobiology Laboratory, Beltsville Agricultural Research Center, USDA-Agricultural Research Service, 20705, Beltsville, MD, USA

    Th. J. Buckhout & Terry C. Hrubec

Authors
  1. Th. J. Buckhout
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Terry C. Hrubec
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

The mention of vendor or product does not imply that they are endorsed or recommended by U.S. Department of Agriculture over vendors of similar products not mentioned.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Buckhout, T.J., Hrubec, T.C. Pyridine nucleotide-dependent ferricyanide reduction associated with isolated plasma membranes of maize (Zea mays L.) roots. Protoplasma 135, 144–154 (1986). https://doi.org/10.1007/BF01277007

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01277007

Keywords

Search

Navigation