Characterization of carbohydrate structural features recognized by anti-arabinogalactan-protein monoclonal antibodies.
Arabinogalactan-proteins (AGPs) are a diverse class of plant cell surface proteoglycans implicated in a range of fundamental processes associated with plant cell development. Anti-AGP monoclonal antibodies have been used extensively for the investigation of the developmental regulation of AGPs although virtually nothing is known about the structure of the carbohydrate epitopes recognised by these antibodies. In this report, a series of methyl glycosides of monosaccharides and a range of oligosaccharides that are elements of the carbohydrate component of AGPs have been investigated for recognition by previously derived anti-AGP monoclonal antibodies. No clear evidence was obtained for the involvement of terminal arabinofuranosides, nor of the galactan backbone, in the recognition of the glycan structure of AGPs by any of the antibodies used in this study. Interestingly, the most effective inhibitor of the binding of the monoclonal antibodies MAC207, JIM4 and JIM13 to exudate gum antigens was an acidic trisaccharide, isolated from a partial acid hydrolysate of gum karaya which has the structure: GlcA beta(1-->3) GalA alpha(1-->2)Rha, determined by a combination of FAB-MS, GC-MS and NMR spectroscopy.
Yates
E A
EA
Centre for Plant Biochemistry & Biotechnology, University of Leeds, United Kingdom.
Valdor
J F
JF
Haslam
S M
SM
Morris
H R
HR
Dell
A
A
Mackie
W
W
Knox
J P
JP
D000911:Antibodies, Monoclonal / Q000276:immunology*; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D002241:Carbohydrates / Q000737:chemistry* / Q000276:immunology; D008401:Gas Chromatography-Mass Spectrometry; D006868:Hydrolysis; D007618:Karaya Gum / Q000737:chemistry; D009682:Magnetic Resonance Spectroscopy; D008969:Molecular Sequence Data; D009088:Mucoproteins / Q000737:chemistry* / Q000276:immunology; D009844:Oligosaccharides / Q000737:chemistry / Q000302:isolation & purification; D010084:Oxidation-Reduction; D010504:Periodic Acid / Q000737:chemistry; D010940:Plant Proteins / Q000737:chemistry* / Q000276:immunology; D012210:Rhamnose / Q000737:chemistry; D016339:Spectrometry, Mass, Fast Atom Bombardment
D000911:Antibodies, Monoclonal; D002241:Carbohydrates; D009088:Mucoproteins; D009844:Oligosaccharides; D010940:Plant Proteins; C006619:arabinogalactan proteins; D010504:Periodic Acid; D007618:Karaya Gum; D012210:Rhamnose