Abstract
The water-insoluble 1,4-β-linked products formed from UDP-[14C]glucose by pea membranes were dissolved in hot dimethyl-sulfoxide/paraformaldehyde and fractionated on columns of controlled pore glass beads calibrated with dextran standards. The products eluted with a peak size close to 70 kilodaltons in dextran equivalents. Similar elution profiles were obtained for products formed in brief or extended incubations and at high or low substrate concentrations. Methylation analysis indicated that only a few [14C]glucose units had been added to an endogenous acceptor to form this product. In the presence of UDP-xylose at concentrations equal to or less than UDP-[14C]glucose, incorporation from the latter was enhanced and the products elongated with time to a size range where the major components eluted between dextran 264 and 500 kilodaltons. Treatment with endo-1,4-β-glucanase resulted in a mixture of oligosaccharides, including the xyloglucan subunit Glc4Xyl3, which were hydrolyzed further by mixed glycosidases to labeled glucose and isoprimeverose (xylosyl-1,6-α-d-glucose). In pulse-chase experiments, the low molecular weight product formed from UDP-[14C]glucose alone was clearly a precursor for high molecular weight products formed subsequently in the presence of both UDP-glucose and UDP-xylose. It is concluded that the 1,4-β-transglucosylation activity detected in these tests was due to an enzyme that is required for biosynthesis of the backbone of xyloglucan.
Full text
PDF
Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Camirand A., Maclachlan G. Biosynthesis of the fucose-containing xyloglucan nonasaccharide by pea microsomal membranes. Plant Physiol. 1986 Oct;82(2):379–383. doi: 10.1104/pp.82.2.379. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Campbell R. E., Brett C. T., Hillman J. R. A xylosyltransferase involved in the synthesis of a protein-associated xyloglucan in suspension-cultured dwarf-French-bean (Phaseolus vulgaris) cells and its interaction with a glucosyltransferase. Biochem J. 1988 Aug 1;253(3):795–800. doi: 10.1042/bj2530795. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hayashi T., Maclachlan G. Pea xyloglucan and cellulose : I. Macromolecular organization. Plant Physiol. 1984 Jul;75(3):596–604. doi: 10.1104/pp.75.3.596. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hayashi T., Matsuda K. Biosynthesis of xyloglucan in suspension-cultured soybean cells. Occurrence and some properties of xyloglucan 4-beta-D-glucosyltransferase and 6-alpha-D-xylosyltransferase. J Biol Chem. 1981 Nov 10;256(21):11117–11122. [PubMed] [Google Scholar]
- Raymond Y., Fincher G. B., Maclachlan G. A. Tissue Slice and Particulate beta-Glucan Synthetase Activities from Pisum Epicotyls. Plant Physiol. 1978 Jun;61(6):938–942. doi: 10.1104/pp.61.6.938. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Saugy M., Farkas V., Maclachlan G. Phosphatases and phosphodiesterases interfere with 1,3-beta-D-glucan synthase activity in pea epicotyl membrane preparations. Eur J Biochem. 1988 Oct 15;177(1):135–138. doi: 10.1111/j.1432-1033.1988.tb14353.x. [DOI] [PubMed] [Google Scholar]
- Spencer F. S., Maclachlan G. A. Changes in Molecular Weight of Cellulose in the Pea Epicotyl during Growth. Plant Physiol. 1972 Jan;49(1):58–63. doi: 10.1104/pp.49.1.58. [DOI] [PMC free article] [PubMed] [Google Scholar]
- TREVELYAN W. E., PROCTER D. P., HARRISON J. S. Detection of sugars on paper chromatograms. Nature. 1950 Sep 9;166(4219):444–445. doi: 10.1038/166444b0. [DOI] [PubMed] [Google Scholar]
- WOOLF L. I. Use of ion-exchange resins in paper chromatography of sugars. Nature. 1953 May 9;171(4358):841–841. doi: 10.1038/171841a0. [DOI] [PubMed] [Google Scholar]