Abundance of Gal beta 1,3GalNAc in O-linked oligosaccharide on hinge region of polymerized IgA1 and heat-aggregated IgA1 from normal human serum.
Gas-phase hydrazinolysis was used to analyze the glycoform of the O-linked oligosaccharide of human serum IgA1. In our previous report, only one glycoform was obtained from the IgA1 of healthy individuals. However, it was found to be composed of heterogeneous IgA1 components having mutually different glycoforms. First, the IgA1 was separated into two subfractions having different affinities toward jacalin. Among them, the high-affinity subfraction was mainly composed of polymerized IgA1. Comparative study of the carbohydrate chain showed a relative abundance of Gal beta 1,3GalNAc in the polymerized form. A simultaneous analysis of the N-glycan of these subfractions was also carried out. Three major components, two biantennary and one triantennary oligosaccharides, were obtained from both subfractions and the relative contents of these components were almost the same. On the other hand, IgA1 was artificially polymerized by heating at 63 degrees C for 2 h. The heat-stable IgA1 was separated from the heat-aggregated material on a Sephacryl S-300 column. The obtained heat-stable IgA1 (approximately 20%) was not further aggregated by more heating under the same conditions. The heat-stable IgA1 contained a much higher amount of the sialylated Gal beta 1,3GalNAc. Thus, it was shown that the degree of completeness of the hinge O-linked oligosaccharide might be correlated with the stability and polymerization process of the IgA1 molecule.
Iwase
H
H
Department of Biochemistry and Medicine, School of Medicine and Nursing, Kitasato University, Sagamihara, Kanagawa.
Tanaka
A
A
Hiki
Y
Y
Kokubo
T
T
Ishii-Karakasa
I
I
Kobayashi
Y
Y
Hotta
K
K
D000116:Acetylgalactosamine / Q000032:analysis; D000915:Antibody Affinity; D001769:Blood / Q000276:immunology*; D002240:Carbohydrate Sequence; D005690:Galactose / Q000032:analysis; D006358:Hot Temperature; D006801:Humans; D007070:Immunoglobulin A / Q000737:chemistry* / Q000302:isolation & purification; D037102:Lectins; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000032:analysis*; D037121:Plant Lectins; D011108:Polymers / Q000737:chemistry*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods
D007070:Immunoglobulin A; D037102:Lectins; D009844:Oligosaccharides; D037121:Plant Lectins; D011108:Polymers; C043696:jacalin; D000116:Acetylgalactosamine; D005690:Galactose