Structural elucidation of N-liked sugar chains of storage glycoproteins in mature pea (Pisum sativum) seeds by ion-spray tandem mass spectrometry (IS-MS/MS).
The structures of N-linked sugar chains of the storage glycoproteins in mature pea seeds have been estimated. Nine pyridylaminated (PA-) N-linked sugar chains were derived and purified from the hydrazinolysate of the storage glycoproteins by reversed-phase HPLC and size-fractionation HPLC. The structures of the PA-sugar chains purified were first identified by two-dimensional PA-sugar chain mapping, considering the data of sugar composition analysis or sequential exoglycosidase digestions. The deduced structures were further analyzed by IS-MS/MS analysis. Every relevant fragment ion derived from all PA-sugar chains could be assigned on the basis of deduced structures. The estimated nine structures fell into two categories; the first was a typical oligomannose type (Man8-3GlcNAc2; 77.7%) which can be hydrolyzed by endo-beta-N-acetylglucosaminidase PS [Y. Kimura et al., Biosci. Biotech. Biochem., 60, 228-232 (1996)], the second was a xylose-containing type (Man4-3Xyl1GlcNAc2, Man3Fuc1Xyl1GlcNAc2; 22.3%). Among these structures, Man8GlcNAc2 (19.7%), Man6GlcNAc2 (24.7%), and Man3Fuc1Xyl1GlcNAc2 (18.8%) were the dominant structures.
Kimura
Y
Y
Department of Bioresources Chemistry, Faculty of Agriculture, Okayama University, Japan.
Ohno
A
A
Takagi
S
S
D002240:Carbohydrate Sequence; D002851:Chromatography, High Pressure Liquid; D006023:Glycoproteins / Q000737:chemistry*; D006026:Glycoside Hydrolases / Q000737:chemistry; D006868:Hydrolysis; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009844:Oligosaccharides / Q000737:chemistry*; D018532:Pisum sativum / Q000737:chemistry*; D012639:Seeds / Q000737:chemistry
D006023:Glycoproteins; D009844:Oligosaccharides; D006026:Glycoside Hydrolases