caa a22 4500 39755740X CHVBK 20180308164806.0 cr unu---uuuuu 161202e199604 xx s 000 0 eng 10.1093/oxfordjournals.jbchem.a021290 doi (NATIONALLICENCE)oxford-10.1093/oxfordjournals.jbchem.a021290 Binding of Anti-Band 3 Autoantibody to Sialylated Poly-N-Acetyllactosaminyl Sugar Chains of Band 3 Glycoprotein on Polyvinylidene Difluoride Membrane and Sepharose Gel: Further Evidence for Anti-Band 3 Autoantibody Binding to the Sugar Chains of Oxidized and Senescent Erythrocytes [Elektronische Daten] [Ken Ando, Klyomi Kikugawa, Masatoshi Beppu] Binding specificity of naturally occurring anti-band 3 IgG antibody isolated from human plasma was investigated in a cell-free binding system. 125I-labeled human anti-band 3 IgG specifically bound to band 3 glycoprotein and lactoferrin, a glycoprotein that has poly-N-acetyllactosamine-type sugar chains like band 3, on the polyvinylidene difluoride blotting membrane. Binding was decreased by 50-70% when band 3 and lactoferrin were pretreated with N-glycosidase F, endo-β-galactosidase, or neuraminidase. Binding of 125I-anti-band 3 IgG to band 3-Sepharose gel was partially inhibited by band 3 oligosaccharides or lactoferrin, but was less inhibited by them after they had been treated with N-glycosidase F or endo-β-galactosidase. A significant part of 125I-anti-band 3 IgG that bound to the band 3-Sepharose gel was released upon treatment of the gel with N-glycosidase F or endo-β-galactosidase. IgG that binds to lactoferrin (anti-lactoferrin IgG) was isolated from normal human plasma. 126I-Anti-lactoferrin IgG bound to the band 3-Sepharose gel as effectively as to the lactoferrin-Sepharose. The antibody specifically bound to the band 3-and lactoferrin-blotted membrane depending on the poly-N-acetyllactosaminyl sugar chains of the blotted glycoproteins. The results indicate that a major part (about 70%) of anti-band 3 IgG recognizes the sialylated poly-N-acetyllactosaminyl sugar chains of band 3 and lactoferrin, and the remaining part (about 30%) of the antibody may recognize the polypeptide portion of band 3. This was supported by the observation that anti-band 3 IgG effectively bound to lactoferrin-Sepharose but 33% of the antibody did not. Anti-band 3 IgG with the carbohydrate-binding property was equally obtained whether fully denatured or barely denatured band 3 was used for isolation of anti-band 3 IgG by affinity chromatog-raphy. These results provide further evidence for our proposal that the binding sites of anti-band 3 IgG to oxidized and senescent erythrocytes reside on the locally condensed sialylated poly-N-acetyllactosaminyl sugar chains of band 3 on the cell surface. © by the Journal of Biochemistry Regular Papers nationallicence anti-band 3 autoantibody nationallicence band 3 glycoprotein nationallicence lactoferrin nationallicence senescent cell antigen nationallicence sialylated poly-N-acetyllactosaminyl sugar chain nationallicence Ando Ken School of Pharmacy, Tokyo University of Pharmacy and Life Science, Hachioji, Tokyo 192-03 aut Kikugawa Klyomi School of Pharmacy, Tokyo University of Pharmacy and Life Science, Hachioji, Tokyo 192-03 aut Beppu Masatoshi School of Pharmacy, Tokyo University of Pharmacy and Life Science, Hachioji, Tokyo 192-03 aut The Journal of Biochemistry Oxford University Press 119/4(1996-04), 639-647 0021-924X 119:4<639 1996 119 jbchem https://doi.org/10.1093/oxfordjournals.jbchem.a021290 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1093/oxfordjournals.jbchem.a021290 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Ando Ken School of Pharmacy, Tokyo University of Pharmacy and Life Science, Hachioji, Tokyo 192-03 aut NATIONALLICENCE 700 1- Kikugawa Klyomi School of Pharmacy, Tokyo University of Pharmacy and Life Science, Hachioji, Tokyo 192-03 aut NATIONALLICENCE 700 1- Beppu Masatoshi School of Pharmacy, Tokyo University of Pharmacy and Life Science, Hachioji, Tokyo 192-03 aut NATIONALLICENCE 773 0- The Journal of Biochemistry Oxford University Press 119/4(1996-04), 639-647 0021-924X 119:4<639 1996 119 jbchem Metadata rights reserved CC BY-NC-4.0 http://creativecommons.org/licenses/by-nc/4.0 nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-oxford