An Arabinogalactan-Protein from Cell Culture of Malva sylvestris

Abstract

An arabinogalactan-protein (AGP) from suspension culture medium of Malva sylvestris was isolated by precipitation with β-glucosyl Yariv reagent, followed by gel-permeation chromatography. It revealed characteristic features of AGPs: a high amount of polysaccharide with a ratio of galactose to arabinose of 1.9 : 1, some uronic acids, and a small protein moiety with the main amino acids serine, alanine and hydroxyproline. The molecular weight was estimated to be 1.3 × 10⁶ Da. Linkage analyses showed that the AGP is composed of a highly branched core polysaccharide of 3-, 6-, and 3,6-linked Galp residues with terminal Araf, GlcAp and Galp. Partial acid hydrolysis resulted in loss of Araf residues at the periphery of the molecule and heavily reduced its reactivity with β-glucosyl Yariv antigen.

Key words

Arabinogalactan proteins - Malva sylvestris - Malvaceae - cell culture - structural analysis - β-glucosyl Yariv reagent

References

• 1 Classen B, Amelunxen F, Blaschek W. Ultrastructural investigations on the development of mucilage idioblasts and cavities of Malva sylvestris ssp. mauritiana .  Scientia Pharmaceutica. 1998;  66 363-80
• 2 Classen B, Blaschek W. High molecular weight acidic polysaccharides from Malva sylvestris and Alcea rosea .  Planta Medica. 1998;  64 640-4
• 3 Bauer R. The Echinacea story - The scientific development of an herbal immunostimulant. In: Prendergast HVD, Etkin NL, Harris DR, Houghton PJ, editors Plants for food and medicine. Royal Botanic Gardens Kew; 1998: 317-32
• 4 Lohmann-Matthes M L, Wagner H. Aktivierung von Makrophagen durch Polysaccharide aus Gewebekulturen von Echinacea purpurea .  Zeitschrift für Phytotherapie. 1989;  10 52-9
• 5 Hoheisel O, Sandberg M, Bertram S, Bulitta M, Schäfer M. Echinagard treatment shortens the course of the common cold: a double-blind, placebo-controlled clinical trial.  European Journal of Clinical Research. 1997;  9 261-8
• 6 Murashige T, Skoog F. A revised medium for rapid growth and bio assays with tobacco tissue cultures.  Physiologia Plantarum. 1962;  15 473-97
• 7 Schumacher D SMZ. personal communication. 
• 8 Yariv J, Rapport M M, Graf L. The interaction of glycosides and saccharides with antibody to the corresponding phenylazo glycosides.  Biochemical Journal. 1962;  85 383-8
• 9 Kreuger M, van Holst G J. Arabinogalactan-protein epitopes in somatic embryogenesis of Daucus carota L.  Planta. 1995;  197 135-41
• 10 Blakeney A B, Harris P J, Henry R J, and Stone B A. A simple and rapid preparation of alditol acetates for monosaccharide analysis.  Carbohydrate Research. 1983;  113 291-9
• 11 Blumenkrantz N, Asboe-Hansen G. New method for quantitative determination of uronic acids.  Analytical Biochemistry. 1973;  54 484-9
• 12 Taylor R L, Conrad H E. Stoichiometric depolymerisation of polyuronides and glycosamino-glyuronans to monosaccharides following reduction of their carbodiimide-activated carboxyl groups.  Biochemistry. 1972;  11 (8) 1383-8
• 13 Harris P J, Henry R J, Blakeney A B, Stone B A. An improved procedure for the methylation analysis of oligosaccharides and polysaccharides.  Carbohydrate Research. 1984;  127 59-73
• 14 Gleeson P A, Clarke A E. Structural studies on the major component of Gladiolus style mucilage, an arabinogalactan-protein.  Biochemical Journal. 1979;  181 607-21
• 15 Jermyn M A, Yeow Y M. A class of lectins present in the tissues of seed plants.  Australian Journal of Plant Physiology. 1975;  2 501-31
• 16 Komalavilas P, Zhu J K, Nothnagel E A. Arabinogalactan-proteins from the suspension culture medium and plasma membrane of rose cells.  Journal of Biological Chemistry. 1991;  266 15 956-65
• 17 Nothnagel E A. Proteoglycans and Related Components in Plant Cells In: International Review of Cytology-A Survey of Cell Biology. Jeon KW, editor Academic Press San Diego; 1997 Vol. 174: 195-291
• 18 Classen B, Blaschek W. Characterization of an arabinogalactan-protein isolated from pressed juice of Echinacea purpurea by precipitation with the β-glucosyl Yariv reagent.  Carbohydrate Research. 2000;  327 497-504
• 19 Idris O HM, Williams P A, Phillips G O. Characterization of gum from Acacia senegal trees of different age and location using multidetection gel permeation chromatography.  Food Hydrocolloids. 1998;  12 379-88
• 20 Aspinall G O. In: Biogenesis of Plant Cell Wall Polysaccharides. Loewus F, editor Academic Press New York; 1973: 95
• 21 Saulnier L, Brillouet J M, Hervé du Penhoat C, Michon V. New investigations of the structure of grape arabinogalactan-protein.  Carbohydrate Research. 1992;  224 219-35

1 * Afterwards vitamins and phytohormones were added as sterilized solutions (filtration).