Association of asialo-galactosyl beta 1-3N-acetylgalactosamine on the hinge with a conformational instability of Jacalin-reactive immunoglobulin A1 in immunoglobulin A nephropathy.
The O-glycans in Jacalin-binding immunoglobulin A1 (IgA1) were released by gas-phase hydrazinolysis and were fractionated by gel filtration and reversed-phase HPLC. Four peaks (P1 to P4) were obtained. There was a significant shift from Peak 2 (monosialylated Gal beta 1-3GalNAc) to Peak 4 (asialo-Gal beta 1-3GalNAc) in the IgA-nephropathy group compared with the negative control group (P < 0.05). One of the functions of the carbohydrate side chains is to stabilize the three-dimensional structure of the glycopeptides. In order to evaluate the stability of the Jacalin-binding IgA1 molecule, the increase in turbidity as a consequence of the increase of the aggregated IgA1 level under the condition of high temperature (63 degrees C) was observed. The increase in the turbidity was significantly higher in the IgA nephropathy group compared with the negative control group (21.7 vs. 5.9% at 150 min, P < 0.02). From a sample of IgA1 solution that had originated from a pooled normal serum, heat-tolerant (nonaggregated) and intolerant (aggregated) IgA1 molecules were separated by gel filtration. The heat-intolerant IgA1 had lower amounts of sialic acid (27.4 micrograms/mg IgA1) than the tolerant IgA1 (37.6 micrograms/mg IgA1). Further, the analysis of the O-glycans released from another IgA1 sample by the hydrazinolysis revealed that the ratio of the asialo-Gal beta 1-3GalNAc/total Gal beta 1-3GalNAc in the heat-in-tolerant IgA1 (27.2%) was increased compared with that in the heat-tolerant IgA1 (18.2%). From these results, it was suggested that unusual glycosylation on the hinge region of Jacalin-binding IgA1 induced an insufficient conformational stiffness to the hinge peptide, resulting in the aggregation of the IgA1 molecule in IgA nephropathy.
Hiki
Y
Y
Department of Medicine, School of Nursing, Kitasato University, Sagamlhara-city, Japan.
Iwase
H
H
Kokubo
T
T
Horii
A
A
Tanaka
A
A
Nishikido
J
J
Hotta
K
K
Kobayashi
Y
Y
D000116:Acetylgalactosamine / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism; D001212:Asialoglycoproteins / Q000737:chemistry* / Q000378:metabolism*; D004355:Drug Stability; D005922:Glomerulonephritis, IGA / Q000378:metabolism*; D006358:Hot Temperature; D006801:Humans; D007070:Immunoglobulin A / Q000737:chemistry / Q000276:immunology*; D037102:Lectins / Q000276:immunology*; D008968:Molecular Conformation; D019158:N-Acetylneuraminic Acid / Q000097:blood; D037121:Plant Lectins; D012016:Reference Values
D001212:Asialoglycoproteins; D007070:Immunoglobulin A; D037102:Lectins; D037121:Plant Lectins; C103104:asialo-galactosyl-acetylgalactosamine; C043696:jacalin; D019158:N-Acetylneuraminic Acid; D000116:Acetylgalactosamine