Isolation of oligomannose-type glycans from bean glycoproteins.
We have isolated individual oligosaccharyl-asparagine derivatives from the total soluble glycoproteins from kidney beens (Phaseolus vulgaris) and from lima beans (Phaseolus limensis). The protein/glycoprotein mixture was digested exhaustively by pronase, and the glycan-containing fractions were separated from free amino acids and peptides by gel filtration. The oligosaccharyl-asparagine derivatives were finally fractionated on Dowex 50 (C. C. Huang, H.E. Meyer, and R. Montgomery, Carbohydr. Res. 13, 127-137, 1970), and the individual fractions were characterized by mass spectrometry, NMR, and ion exchange chromatography. With the procedures described, only oligomannose derivatives were obtained from the beans. In the case of kidney beans, six different derivatives were observed and characterized, Man9GlcNAc2Asn, two positional isomers of Man8GlcNAc2Asn, two positional isomers of Man7GlcNAc2Asn, and Man6GlcNAc2Asn. Under identical conditions the lima beans yielded primarily the Man9GlcNAc2Asn derivative along with a small amount of the two Man8GlcNAc2Asn derivatives. The oligomannose structures can be isolated in reasonable quantities (2-20 mg) from about 200 g of dry beans.
Lu
Y
Y
Department of Biochemistry and Molecular Biology, University of Texas Medical School, Houston 77225.
Ye
J
J
Wold
F
F
D001216:Asparagine / Q000032:analysis; D002240:Carbohydrate Sequence; D007887:Fabaceae / Q000737:chemistry* / Q000201:enzymology; D006023:Glycoproteins / Q000032:analysis*; D008358:Mannose / Q000302:isolation & purification*; D008361:Mannosidases / Q000378:metabolism; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000302:isolation & purification*; D010940:Plant Proteins / Q000032:analysis*; D010946:Plants, Medicinal
D006023:Glycoproteins; D009844:Oligosaccharides; D010940:Plant Proteins; D001216:Asparagine; D008361:Mannosidases; D008358:Mannose