Amano J, Straehl P, Berger E G, Kochibe N, Kobata A
Department of Biochemistry, University of Tokyo, Japan.
J Biol Chem. 1991 Jun 25;266(18):11461-77.
The mucin-type sugar chains of human milk galactosyltransferase samples purified from two donors with different blood types were released by alkaline borohydride treatment and quantitatively labeled by N-[3H]acetylation. The radioactive oligosaccharides thus obtained were fractionated by high performance liquid chromatography and immobilized lectin chromatography, and their structures were studied by sequential digestion with endo- or exoglycosidases, methylation analysis, and periodate oxidation. It was revealed that the structures of the mucin-type sugar chains of galactosyltransferase are extremely various, and many blood group determinants are expressed on more than 13 different backbone sugar chains. The characteristic features of the sugar chains could be summarized as follows. 1) The sugar chains of both samples are composed of core 1, Gal beta 1----3GalNAc, and core 2, GlcNAc beta 1----6(Gal beta 1----3)GalNAc. 2) One or two N-acetyllactosamine repeating units extend from the core through GlcNAc beta 1----6Gal and GlcNAc beta 1----3 Gal linkages. 3) Blood group determinants are expressed in accord with the blood types of the donors: sample 1 from a donor of blood type O, Lea+b- contains oligosaccharides with Lea and X determinants, and sample 2 from a donor of B, Lea-b- contains those with H, X, Y, and B determinants.
从两名不同血型的供体中纯化得到的人乳半乳糖基转移酶样品的粘蛋白型糖链,经碱性硼氢化物处理后释放,并通过N-[³H]乙酰化进行定量标记。由此获得的放射性寡糖通过高效液相色谱和固定化凝集素色谱进行分离,其结构通过内切或外切糖苷酶的顺序消化、甲基化分析和高碘酸盐氧化进行研究。结果表明,半乳糖基转移酶的粘蛋白型糖链结构极其多样,并且在超过13种不同的主链糖链上表达了许多血型决定簇。糖链的特征可总结如下。1)两个样品的糖链均由核心1(Galβ1----3GalNAc)和核心2(GlcNAcβ1----6(Galβ1----3)GalNAc)组成。2)一个或两个N-乙酰乳糖胺重复单元通过GlcNAcβ1----6Gal和GlcNAcβ1----3Gal连接从核心延伸。3)血型决定簇根据供体的血型表达:来自O型血供体的样品1,Lea+b-含有带有Lea和X决定簇的寡糖,来自B型血供体的样品2,Lea-b-含有带有H、X、Y和B决定簇的寡糖。