Tanner M J, Anstee D J, Mallinson G, Ridgwell K, Martin P G, Avent N D, Parsons S F
Department of Biochemistry, University of Bristol, United Kingdom.
Carbohydr Res. 1988 Jul 15;178:203-12. doi: 10.1016/0008-6215(88)80112-5.
Endo-N-acetyl-beta-D-glucosaminidase F-Peptidyl N-glycosidase F preparations (abbreviated Endo F) and endo-beta-D-galactosidase were used to study the major human erythrocyte membrane glycoproteins and the components carrying the blood group A, B, Rhesus (D), and Duffy (Fya) antigens. The results are consistent with the known presence of an N-glycosyl-linked oligosaccharide on sialoglycoprotein alpha and the absence of such an oligosaccharide from sialoglycoprotein delta. Under the conditions used, only a portion of the N-glycosyl-linked oligosaccharides on band 3 molecules were cleaved by Endo F alone or by Endo F in combination with endo-beta-D-galactosidase. Immunoblotting experiments showed that treatment of red cells with Endo F alone had little effect on the components carrying blood group A and B antigen activity. However, Endo F used in combination with endo-beta-D-galactosidase caused a substantial reduction in the binding of monoclonal anti-A and anti-B antibodies. The results clearly show that sialoglycoproteins alpha and delta carry little or no blood group A or B activity. Endo F alone, or in combination with endo-beta-D-galactosidase, had no effect on the electrophoretic mobility of the Rh(D) polypeptide, supporting previous suggestions that this membrane polypeptide is unusual in not being glycosylated. Endo F had a dramatic effect on the electrophoretic mobility of the component(s) carrying blood group Fya activity. The diffuse Fya component of Mr 38,500-90,000 was sharpened to a band of Mr 26,000. Either endo-beta-D-galactosidase or neuraminidase treatment reduced the Mr of the Fya component(s) but did not significantly sharpen the bands, suggesting that the Fya component contains between 40-50% by mass of N-glycosyl-linked oligosaccharides.
内切-N-乙酰-β-D-氨基葡萄糖苷酶F-肽基N-糖苷酶F制剂（简称为内切F）和内切-β-D-半乳糖苷酶被用于研究主要的人类红细胞膜糖蛋白以及携带A、B、Rh（D）和达菲（Fya）血型抗原的成分。结果与已知的唾液酸糖蛋白α上存在N-糖基连接寡糖以及唾液酸糖蛋白δ上不存在这种寡糖的情况一致。在所使用的条件下，仅一部分带3分子上的N-糖基连接寡糖可被单独的内切F或内切F与内切-β-D-半乳糖苷酶联合切割。免疫印迹实验表明，单独用内切F处理红细胞对携带A和B血型抗原活性的成分影响很小。然而，内切F与内切-β-D-半乳糖苷酶联合使用会导致单克隆抗A和抗B抗体的结合显著减少。结果清楚地表明，唾液酸糖蛋白α和δ几乎不携带或不携带A或B血型活性。单独的内切F或与内切-β-D-半乳糖苷酶联合使用，对Rh（D）多肽的电泳迁移率没有影响，这支持了之前关于这种膜多肽未被糖基化的说法。内切F对携带Fya血型活性的成分的电泳迁移率有显著影响。38,500 - 90,000 Mr的弥散性Fya成分锐化成为一条26,000 Mr的条带。内切-β-D-半乳糖苷酶或神经氨酸酶处理均可降低Fya成分的Mr，但并未显著锐化条带，这表明Fya成分含有40 - 50%质量的N-糖基连接寡糖。
