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Immunochemical comparison of membrane-associated and secreted arabinogalactan-proteins in rice and carrot

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Abstract

Arabinogalactan-proteins (AGPs) occurring in suspension-cultured rice (Oryza saliva L.) cells, their conditioned medium and at the rice root apex were investigated using monoclonal antibodies and the AGP-binding β-glucosyl Yariv reagent (β GlcY). A monoclonal antibody, LM2, was generated that recognized an acidic carbohydrate epitope common to two soluble AGPs occurring in the conditioned medium of proliferating rice cells, membrane-associated AGPs (rmAGP) in the cultured cells and two AGPs at the rice root apex. In addition, LM2 recognized AGPs secreted by suspensioncultured carrot (Daucus carota L.) cells. The two AGPs of the rice culture medium, srAGP1 and srAGP2, were discriminated by their mobilities during sodium dodecyl sulfate-polyacrylamide gel electrophoresis, reaction with β GlcY, the presence of arabinogalactan epitopes and anion-exchange chromatography. The association of rmAGP with the plasma membrane was investigated by Triton-X-114/aqueous partitioning of both microsomal and plasma-membrane preparations and rmAGP was found to partition into the detergent phase, indicating that AGPs are hydrophobic plasma-membrane proteins in rice. This was in contrast to plasma-membrane AGPs of suspension-cultured carrot cells that partitioned into the aqueous phase. At the rice root apex most of the AGP was associated with the microsomal fraction and also partitioned into the detergent phase, although a distinct highmolecular-mass AGP entered the aqueous phase.

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Abbreviations

AGP:

arabinogalactan-protein

βGlcY:

β-glucosyl Yariv reagent

ELISA:

enzyme-linked immunosorbent assay

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Author notes

  1. Margaret Smallwood

    Present address: The Plant Laboratory, Department of Biology, University of York, Y01 5YW, York, UK

  2. Edwin A. Yates

    Present address: Istituto Scientifico di Chimica e Biochimica “G. Ronzoni”, Via G, Colombo 81, 1-20133, Milano, Italy

Authors and Affiliations

  1. Centre for Plant Biochemistry & Biotechnology, University of Leeds, LS2 9JT, Leeds, UK

    Margaret Smallwood, Edwin A. Yates, William G. T. Willats, Helen Martin & J. Paul Knox

Authors
  1. Margaret Smallwood
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  2. Edwin A. Yates
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  3. William G. T. Willats
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  4. Helen Martin
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  5. J. Paul Knox
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Additional information

We gratefully acknowledge support from the Leverhulme Trust, the UK Biotechnology and Biological Sciences Research Council and the Royal Society.

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Smallwood, M., Yates, E.A., Willats, W.G.T. et al. Immunochemical comparison of membrane-associated and secreted arabinogalactan-proteins in rice and carrot. Planta 198, 452–459 (1996). https://doi.org/10.1007/BF00620063

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  • DOI: https://doi.org/10.1007/BF00620063

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