Synthesis of (1-->3), (1-->4)-beta-D-glucan in the Golgi apparatus of maize coleoptiles
| Autoři: | David M. Gibeaut, Nicholas C. Carpita |
|---|---|
| Zdroj: | Proceedings of the National Academy of Sciences. 90:3850-3854 |
| Informace o vydavateli: | Proceedings of the National Academy of Sciences, 1993. |
| Rok vydání: | 1993 |
| Témata: | Chromatography, Gas, Sucrose, Molecular Sequence Data, Golgi Apparatus, Oligosaccharides, Biology, Polysaccharide, Zea mays, chemistry.chemical_compound, symbols.namesake, Cellulase, Cell Wall, Dextrins, Carbohydrate Conformation, Tetrasaccharide, Trisaccharide, Cellulose, Glucans, Chromatography, High Pressure Liquid, Glucan, chemistry.chemical_classification, Multidisciplinary, Chromatography, Callose, Golgi apparatus, carbohydrates (lipids), Carbohydrate Sequence, chemistry, Biochemistry, Glucosyltransferases, Chromatography, Gel, symbols, Research Article, Homogenization (biology) |
| Popis: | Membranes of the Golgi apparatus from maize (Zea mays L.) were used to synthesize in vitro the (1-->3), (1-->4)-beta-D-glucan (MG) that is unique to the cell wall of the Poaceae. The MG was about 250 kDa and was separated from a much larger (1-->3)-beta-D-glucan (callose) by gel-permeation chromatography. Diagnostic oligosaccharides, released by a sequence-dependent endoglucanase from Bacillus subtilis, were separated by HPLC and GLC. The trisaccharide beta-D-Glcp-(1-->4)-beta-D-Glcp-(1-->3)-D-Glc, the tetrasaccharide [beta-D-Glcp-(1-->4)]2-beta-D-Glcp-(1-->3)-D-Glc, and longer cellodextrin-(1-->3)-D-Glc oligosaccharides were synthesized in proportions similar to those found in purified MG. Activated charcoal added during homogenization enhanced synthesis of MG, presumably by removing inhibitory compounds. The Golgi apparatus was determined as the site of synthesis by a combination of downward and flotation centrifugations on sucrose step gradients. The rate of synthesis did not reach saturation at up to 10 mM UDP-Glc. Chelators completely abolished synthesis, but synthase activity was restored by addition of either MgCl2 or, to a lesser extent, MnCl2. Synthesis continued for well over 1 h; addition of KOH to raise the pH from 7.2 to 8.0 during the reaction increased the rate of synthesis, which indicates that a transmembrane pH gradient may facilitate synthesis of MG. |
| ISSN: | 1091-6490 0027-8424 |
| DOI: | 10.1073/pnas.90.9.3850 |
| Přístupová URL adresa: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::4a9be6574dfe8c5a44b1bcaefc2880f9 https://doi.org/10.1073/pnas.90.9.3850 |
| Rights: | OPEN |
| Přírůstkové číslo: | edsair.doi.dedup.....4a9be6574dfe8c5a44b1bcaefc2880f9 |
| Autor: | David M. Gibeaut, Nicholas C. Carpita |
| Rok vydání: | 1993 |
| Předmět: |
Chromatography
Gas Sucrose Molecular Sequence Data Golgi Apparatus Oligosaccharides Biology Polysaccharide Zea mays chemistry.chemical_compound symbols.namesake Cellulase Cell Wall Dextrins Carbohydrate Conformation Tetrasaccharide Trisaccharide Cellulose Glucans Chromatography High Pressure Liquid Glucan chemistry.chemical_classification Multidisciplinary Chromatography Callose Golgi apparatus carbohydrates (lipids) Carbohydrate Sequence chemistry Biochemistry Glucosyltransferases Chromatography Gel symbols Research Article Homogenization (biology) |
| Zdroj: | Proceedings of the National Academy of Sciences. 90:3850-3854 |
| ISSN: | 1091-6490 0027-8424 |
| DOI: | 10.1073/pnas.90.9.3850 |
| Popis: | Membranes of the Golgi apparatus from maize (Zea mays L.) were used to synthesize in vitro the (1-->3), (1-->4)-beta-D-glucan (MG) that is unique to the cell wall of the Poaceae. The MG was about 250 kDa and was separated from a much larger (1-->3)-beta-D-glucan (callose) by gel-permeation chromatography. Diagnostic oligosaccharides, released by a sequence-dependent endoglucanase from Bacillus subtilis, were separated by HPLC and GLC. The trisaccharide beta-D-Glcp-(1-->4)-beta-D-Glcp-(1-->3)-D-Glc, the tetrasaccharide [beta-D-Glcp-(1-->4)]2-beta-D-Glcp-(1-->3)-D-Glc, and longer cellodextrin-(1-->3)-D-Glc oligosaccharides were synthesized in proportions similar to those found in purified MG. Activated charcoal added during homogenization enhanced synthesis of MG, presumably by removing inhibitory compounds. The Golgi apparatus was determined as the site of synthesis by a combination of downward and flotation centrifugations on sucrose step gradients. The rate of synthesis did not reach saturation at up to 10 mM UDP-Glc. Chelators completely abolished synthesis, but synthase activity was restored by addition of either MgCl2 or, to a lesser extent, MnCl2. Synthesis continued for well over 1 h; addition of KOH to raise the pH from 7.2 to 8.0 during the reaction increased the rate of synthesis, which indicates that a transmembrane pH gradient may facilitate synthesis of MG. |
| Databáze: | OpenAIRE |
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