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Characterization of an arabinogalactan-protein from suspension culture of Echinacea purpurea

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Abstract

Suspension cultures of Echinacea purpurea have been established in MS medium supplemented with 2,4-D and an arabinogalactan-protein (AGP) was purified from the secreted soluble polymers by precipitation with ethanol, followed by precipitation with β-glucosyl Yariv reagent. It revealed typical features of AGPs: a high amount of polysaccharide (90% w/w) with the dominating monosaccharides galactose and arabinose and some glucuronic acid, and a small protein moiety (10% w/w) with the main amino acids Ala, Hyp, Glx, Ser, Asx and Thr. Linkage- and NMR-analyses showed the polysaccharide part to be composed of a branched core-polysaccharide of 3-, 6- and 3,6-linked Galp residues with terminal Araf, Arap, Galp and GlcAp residues. Compared to an AGP from pressed juice of the aerial parts of Echinacea purpurea, differences particularly in terminal arabinose mono- and oligosaccharides in arabinogalactan (AG) side branches could be detected. Testing of different AGP-antibodies with both AGPs confirmed the results of the analytical investigations. Binding of AGPs from plant and cell cultures to LM2, a monoclonal AGP-antibody reacting with a GlcA containing epitope, was comparable. The reactivity of a monoclonal antibody raised against the AGP from the plant recognizing a galactan epitope was also nearly similar with both AGPs. In contrast, polyclonal antibodies raised against the AGP from the plant and directed against an Araf-containing epitope of the AG side branches showed nearly no cross reactivity with the AGP from cell culture.

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Abbreviations

AG:

Arabinogalactan

AGP:

Arabinogalactan-protein

2,4-D:

2,4-Dichlorophenoxyacetic acid

GLC–MS:

Gas liquid chromatography–mass spectrometry

MS:

Murashige and Skoog medium (1962)

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Acknowledgements

Thanks to F. Rodde for technical assistance, to Dr. I. Zündorf, Frankfurt, Germany, for generation of the monoclonal antibody against Echinacea AGP and to Prof. P. Knox, Leeds, and Prof. K. Roberts, Norwich, UK, for providing LM 2.

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  1. Institute of Pharmacy, Department of Pharmaceutical Biology, University of Kiel, Gutenbergstr. 76, 24118, Kiel, Germany

    Birgit Classen

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  1. Birgit Classen
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Correspondence to Birgit Classen.

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Classen, B. Characterization of an arabinogalactan-protein from suspension culture of Echinacea purpurea . Plant Cell Tiss Organ Cult 88, 267–275 (2007). https://doi.org/10.1007/s11240-006-9197-9

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  • DOI: https://doi.org/10.1007/s11240-006-9197-9

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