l-Galactose replaces l-fucose in the pectic polysaccharide rhamnogalacturonan II synthesized by the l-fucose-deficient mur1 Arabidopsis mutant

Arabidopsis thaliana mur1 is a dwarf mutant with altered cell-wall properties, in which l-fucose is partially replaced by l-galactose in the xyloglucan and glycoproteins. We found that the mur1 mutation also affects the primary structure of the pectic polysaccharide rhamnogalacturonan II (RG-II). In mur1 RG-II a non-reducing terminal 2-O-methyl l-galactosyl residue and a 3,4-linked l-galactosyl residue replace the non-reducing terminal 2-O-methyl l-fucosyl residue and the 3,4-linked l-fucosyl residue, respectively, that are present in wild-type RG-II. Furthermore, we found that a terminal non-reducing l-galactosyl residue, rather than the previously reported d-galactosyl residue, is present on the 2-O-methyl xylose-containing side chain of RG-II in both wild type and mur1 plants. Approximately 95% of the RG-II from wild type and mur1 plants is solubilized as a high-molecular-weight (>100 kDa) complex, by treating walls with aqueous potassium phosphate. The molecular mass of RG-II in this complex was reduced to 5–10 kDa by treatment with endopolygalacturonase, providing additional evidence that RG-II is covalently linked to homogalacturonan. The results of this study provide additional information on the structure of RG-II and the role of this pectic polysaccharide in the plant cell wall.Abbreviations AIR Alcohol-insoluble residue - d-Gal d-Galactosyl - EPG Endopolygalacturonase - ESI–MS Electrospray ionization mass spectrometry - GC–MS Gas chromatography–mass spectrometry - ¹H-NMR Proton nuclear magnetic resonance spectroscopy - l-Fuc l-Fucosyl - l-Gal l-Galactosyl - 2-O-MeFuc 2-O-Methyl l-fucosyl - 2-O-MeGal 2-O-Methyl l-galactosyl - 2-O-MeXyl 2-O-Methyl d-xylosyl - MWCO Molecular weight cut-off - RG-II Rhamnogalacturonan II - ppm Parts per million - RI Refractive index - SEC Size-exclusion chromatography - TFA Trifluoroacetic acid - WT Wild type