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J Biol Chem, Vol. 274, Issue 19, 13098-13104, May 7, 1999

The Plant Cell Wall Polysaccharide Rhamnogalacturonan II Self-assembles into a Covalently Cross-linked Dimer

Tadashi IshiiDagger , Toshiro Matsunaga§, Patrice Pellerin, Malcolm A. O'Neillparallel , Alan Darvillparallel , and Peter Albersheimparallel

From the Dagger  Forestry and Forest Products Research Institute, Tsukuba Norin Kenkyu, Danchi-Nai, Ibaraki 305-8687, Japan, the § Kyushu National Agricultural Experiment Station, Nishigoshi, Kumamoto 861-1192, Japan, the  Institut National de la Recherche Agronomique, Institut des Produits de la Vigne, Unite de Recherches Biopolymeres et Aromes, 2 Place Viala, F-34060 Montpellier cedex, France, and the parallel  Complex Carbohydrate Research Center and Department of Biochemistry and Molecular Biology, the University of Georgia, Athens, Georgia 30602-4712

The location of the 1:2 borate-diol ester cross-link in the dimer of the plant cell wall polysaccharide rhamnogalacturonan II (RG-II) has been determined. The ester cross-links the apiofuranosyl residue of the 2-O-methyl-D-xylose-containing side chains in each of the subunits of the dimer. The apiofuranosyl residue in each of the two aceric acid-containing side chains is not esterified. The site of borate esterification is identical in naturally occurring and in in vitro synthesized dimer. Pb2+, La3+, and Ca2+ increase dimer formation in vitro in a concentration- and pH-dependent manner. Pb2+ is the most effective cation. The dimer accounts for 55% of the RG-II when the monomer (0.5 mM) is treated for 5 min at pH 3.5 with boric acid (1 mM) and Pb2+ (0.5 mM); at pH 5 the rate of conversion is somewhat slower. Hg2+ does not increase the rate of dimer formation. A cation's charge density and its ability to form a coordination complex with RG-II, in addition to steric factors, may regulate the rate and stability of dimer formation in vitro. Our data provide evidence that the structure of RG-II itself determines which apiofuranosyl residues are esterified with borate and that in the presence of boric acid and certain cations, two RG-II monomers self-assemble to form a dimer.

Copyright © 1999 by The American Society for Biochemistry and Molecular Biology, Inc.


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