促红细胞生成素中硫酸化 N-连接寡糖的结构分析
Structural analysis of sulfated N-linked oligosaccharides in erythropoietin.
作者信息
Kawasaki N, Haishima Y, Ohta M, Itoh S, Hyuga M, Hyuga S, Hayakawa T
机构信息
Division of Biological Chemistry and Biologicals, National Institute of Health Sciences, 1-18-1, Kamiyoga, Setagaya-ku, Tokyo 158-8501 Japan.
出版信息
Glycobiology. 2001 Dec;11(12):1043-9. doi: 10.1093/glycob/11.12.1043.
We previously demonstrated that high-performance liquid chromatography with electrospray ionization mass spectrometry (LC/MS) equipped with a graphitized carbon column (GCC) is useful for the structural analysis of carbohydrates in glycoproteins. Using LC/MS with GCC, sulfated N-linked oligosaccharides were found in erythropoietin (EPO) expressed in baby hamster kidney cells. Sulfation occurs in a part of the N-linked oligosaccharides in the EPO. Sulfated monosaccharide residue in the sulfated N-linked oligosaccharide was determined by exoglycosidase digestion followed by sugar mapping by LC/MS. The linkage position and branch-location of the sulfate group in the tetraantennary oligosaccharide were analyzed by (1)H-nuclear magnetic resonance. It was suggested that sulfation occurs on the C-6 position of GlcNAc located in the GlcNAcbeta1-4Manalpha1-3 branch.
我们之前证明，配备石墨化碳柱（GCC）的高效液相色谱-电喷雾电离质谱联用仪（LC/MS）可用于糖蛋白中碳水化合物的结构分析。使用配备GCC的LC/MS，在幼仓鼠肾细胞表达的促红细胞生成素（EPO）中发现了硫酸化的N-连接寡糖。硫酸化发生在EPO中一部分N-连接寡糖上。通过外切糖苷酶消化，然后用LC/MS进行糖图谱分析，确定了硫酸化N-连接寡糖中的硫酸化单糖残基。通过¹H-核磁共振分析了四天线寡糖中硫酸基团的连接位置和分支位置。结果表明，硫酸化发生在位于GlcNAcbeta1-4Manalpha1-3分支中的GlcNAc的C-6位上。
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