Skip to main content
SpringerLink
Log in

Cytoskeleton components of inside-out and right-side-out plasma membrane vesicles from plants

  • Published:
Protoplasma Aims and scope Submit manuscript

Summary

Isolated plasma membrane vesicles purified by aqueous polymer two-phase partitioning were used as a model system for studies on the membrane-associated (cortical) cytoskeleton in plants. Actin, as identified by immunoblotting, was found to be specifically attached to plasma membrane vesicles from cauliflower (Brassica oleracea L.). The actin was not washed off as the vesicles were turned inside-out, indicative of a fairly strong attachment. Triton X-100 extraction of plasma membrane vesicles resulted in an insoluble and hence pelletable fraction where actin could be found together with several other proteins. Our results show that the cortical cytoskeleton is to some extent co-purified with the plasma membrane, and we believe that well defined, inside-out and right-side-out plasma membrane vesicles can be used to study the structure and dynamics of the plant cortical cytoskeleton.

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

ATP:

adenosine 5′-triphosphate

BCIP:

5-bromo-4-chloro-3-indolyl phosphate

BSA:

bovine serum albumin

CCD:

counter-current distribution

DTT:

dithiothreitol

EDTA:

ethylene-diamine-tetraacetic acid

EGTA:

ethylene glycol-bis(β-aminoethyl ether)

GSII:

1,3-β-glucan synthase

HEPES:

N-[2-hydroxyethyl]-piperazine-N′-[2-ethane sulfonic acid]

MES:

2-(N-morpholino)ethane sulfonic acid

NBT:

p-nitro blue tetrazolium chloride

IDP:

inosine 5′-diphosphate

PAGE:

polyacrylamide gel electrophoresis

PBS:

phosphate buffered saline

PIPES:

piperazine-N,N′-bis(2-ethane-sulfonic acid)

PPB:

potassium phosphate buffer

PM:

plasma membrane

PMSF:

phenylmethylsulfonyl fluoride

PVDF:

polyvinylidene difluoride

PVPP:

polyvinylpolypyrrolidone

SDS:

sodium dodecyl sulfate

TBS:

Tris-buffered saline

TTBS:

Tris-buffered saline with Tween 20

Tris:

tris(hydroxymethyl) aminomethane

References

  • Abe S, Ito Y, Davies E (1992) Co-sedimentation of actin, tubulin and membranes in the cytoskeleton fractions from peas and mouse 3T3 cells. J Exp Bot 43: 941–949

    Google Scholar 

  • Askerlund P (1990) Redox processes of plant plasma membranes. Studies with isolated plasma membrane vesicles. PhD Thesis, University of Lund, Lund, Sweden

    Google Scholar 

  • —, Larsson C, Widell S (1988) Localizaton of donor and acceptor sites of dehydrogenase activities using inside-out and right-sideout plasma membrane vesicles from plants. FEBS Lett 239: 23–28

    Google Scholar 

  • Baginski ES, Foa PP, Zak B (1967) Determination of phosphate: study of labile organic phosphate interference. Clin Chim Acta 15: 155–158

    Google Scholar 

  • Bearden JC Jr (1978) Quantification of submicrogram quantities of protein by an improved protein-dye binding assay. Biochim Biophys Acta 533: 525–529

    PubMed  Google Scholar 

  • Berridge MJ, Irvine RF (1989) Inositol phosphates and cell signalling. Nature 341: 197–205

    PubMed  Google Scholar 

  • Drøbak BK (1991) Plant signal perception and transduction: the role of the phosphoinositide system. Essays Biochem 26: 27–37

    PubMed  Google Scholar 

  • Faraday CD, Spanswick R (1993) Evidence for a membrane skeleton in higher plants. A spectrin-like polypeptide co-isolates with rice root plasma membranes. FEBS Lett 318: 313–316

    PubMed  Google Scholar 

  • Fredrikson K, Larsson C (1989) Activation of 1,3-β-glucan synthase by Ca2 +, spermine and cellobiose. Localization of activator sites using inside-out plasma membrane vesicles. Physiol Plant 77: 196–201

    Google Scholar 

  • Haupt W, Scheuerlein R (1990) Chloroplast movement. Plant Cell Environ 13: 595–614

    Google Scholar 

  • Hepler PK, Wayne RO (1985) Calcium and plant development. Annu Rev Plant Physiol 36: 397–439

    Google Scholar 

  • Kropf DL, Berge SK, Quatrano RS (1989) Actin localization duringFucus embryogenesis. Plant Cell 1: 191–200

    PubMed  Google Scholar 

  • Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685

    PubMed  Google Scholar 

  • Larsson C (1985) Plasma membranes. In: Linskens HF, Jackson JF (eds) Cell components. Springer, Berlin Heidelberg New York Tokyo, pp 85–104 (Modern methods of plant analysis, new series, vol 1)

    Google Scholar 

  • —, Kjellbom P, Widell S, Lundborg T (1984) Sidedness of plant plasma membrane vesicles purified by partitioning in aqueous two-phase systems. FEBS Lett 171: 271–276

    Google Scholar 

  • —, Widell S, Kjellbom P (1987) Preparation of high-purity plasma membranes. Methods Enzymol 148: 558–568

    Google Scholar 

  • Lassing I, Lindberg U (1985) Specific interactions between phosphatidylinositol 4,5-bisphosphate and profilactin. Nature 314: 472–474

    PubMed  Google Scholar 

  • Lundborg T, Widell S, Larsson C (1981) Distribution of ATPases in wheat root membranes separated by phase partition. Physiol Plant 52: 89–95

    Google Scholar 

  • Mescher MF, Jose MJL, Balk SP (1981) Actin containing matrix associated with the plasma membrane of murine tumour and lymphoid cells. Nature 289: 139–144

    PubMed  Google Scholar 

  • Niggli V, Jenni V (1989) Actin-associated proteins in human neutrophils: identification and reorganization upon cell activation. Eur J Cell Biol 49: 366–372

    PubMed  Google Scholar 

  • Palmgren MG, Sommarin M (1989) Lysophosphatidylcholine stimulates ATP dependent proton accumulation in isolated root plasma membrane vesicles. Plant Physiol 90: 1009–1014

    Google Scholar 

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

    Google Scholar 

  • —, Sommarin M, Ulvskov P, Larsson C (1990 b) Effect of detergents on the proton ATPase activity of inside-out and right-side-out plant plasma membrane vesicles. Biochim Biophys Acta 1021: 133–140

    PubMed  Google Scholar 

  • Payrastre B, van Bergen EN, Henegouwen PMP, Breton M, den Hartigh JC, Plantavid M, Verkleij AJ, Boonstra J (1991) Phosphoinositide kinase, diacylglycerol kinase, and phospholipase C activities associated to the cytoskeleton: effect of epidermal growth factor. J Cell Biol 115: 121–128

    PubMed  Google Scholar 

  • Sandelius AS, Morré DJ (1990) Plasma membrane isolation. In: Larsson C, Møller IM (eds) The plant plasma membrane. Springer, Berlin Heidelberg New York Tokyo, pp 44–75

    Google Scholar 

  • Schliwa M, Blerkom J (1990) Structural interaction of cytoskeletal components. J Cell Biol 90: 222–235

    Google Scholar 

  • Seitz K (1979) Cytoplasmic streaming and cyclosis of chloroplasts. In: Haupt W, Feinleib ME (eds) Physiology of movements. Springer, Berlin Heidelberg New York, pp 150–169 [Pirson A, Zimmermann MH (eds) Encyclopedia of plant physiology, new series, vol 7]

    Google Scholar 

  • Shariff A, Luna EJ (1992) Diacylglycerol-stimulated formation of actin nucleation sites at plasma membranes. Science 256: 245–247

    PubMed  Google Scholar 

  • Sommarin M, Sandelius AS (1988) Phosphatidylinositol and phosphatidylinositol phosphate kinase in plant plasma membranes. Biochim Biophys Acta 958: 268–278

    Google Scholar 

  • Spudich JA, Watt S (1971) The regulation of rabbit skeletal muscle contraction. J Biol Chem 246: 4866–4871

    PubMed  Google Scholar 

  • Traas JA (1990) The plasma membrane-associated cytoskeleton. In: Larsson C, Møller IM (eds) The plant plasma membrane. Springer, Berlin Heidelberg New York Tokyo, pp 269–292

    Google Scholar 

  • Tan Z, Boss W (1992) Association of phosphatidylinositol kinase, phosphatidylinositol monophosphate kinase and diacylglycerol kinase with the cytoskeleton and F-actin fractions of carrot (Daucus carota L.) cells grown in suspension culture. Plant Physiol 100: 2116–2120

    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 

  • —, Sommarin M (1991) Purification of endoplasmic reticulum from wheat roots and shoots (Triticum aestivum). Comparison of their blue light-sensitive redox components with those of highly purified plasma membrane. Physiol Plant 82: 9–18

    Google Scholar 

  • Williamson RE, McCurdy DW, Hurley UA, Perkin JL (1987) Actin ofChara giant internodal cells. A single isoform in the subcortical filament bundles and a larger, immunologically related protein in the chloroplasts. Plant Physiol 85: 268–272

    Google Scholar 

  • Xu P, Lloyd CW, Staiger CJ, Drøbak BK (1992) Association of phosphatidylinositol 4-kinase with the plant cytoskeleton. Plant Cell 4: 941–951

    PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Plant Biology, Section of Plant Physiology, University of Lund, P.O. Box 7007, S-220 07, Lund, Sweden

    A. Sonesson & Susanne Widell

Authors
  1. A. Sonesson
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Susanne Widell
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Sonesson, A., Widell, S. Cytoskeleton components of inside-out and right-side-out plasma membrane vesicles from plants. Protoplasma 177, 45–52 (1993). https://doi.org/10.1007/BF01403398

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01403398

Keywords

Search

Navigation