van Rooijen J J, Jeschke U, Kamerling J P, Vliegenthart J F
Bijvoet Center, Department of Bio-Organic Chemistry, Utrecht University, P.O. Box 80075, NL-3508 TB Utrecht, The Netherlands, Universität Rostock, Frauenklinik, P.O. Box 18055, Rostock, Germany.
Glycobiology. 1998 Nov;8(11):1053-64. doi: 10.1093/glycob/8.11.1053.
Transferrin, a glycoprotein involved in iron transport in body fluids, was isolated from amniotic fluid of a hydramniospatient by sequential anion-exchange chromatography and gel filtration. The N-glycans of human amniotic fluid transferrin (hAFT) were enzymatically liberated by PNGase-F digestion, isolated by gel filtration and fractionated by (high-pH) anion-exchange chromatography. After alkaline borohydride treatment of native hAFT, the released O-glycans were isolated by gel filtration and fractionated by anion-exchange chroma-tography. Structure elucidation of 14 N- and 2 O-glycans was performed by 500 or 600 MHz1H-NMR spectroscopy. Besides conventional N-glycans established earlier for human serum transferrin (hST), new (alpha1-3)-fucosylated N-glycans were found, representing sialyl Le(x) elements. Furthermore, as compared to hST, a higher degree of (alpha1-6)-fucosylation and an increase in branching from di- to triantennary compounds has been detected. The presence of O-glycans is demonstrated for the first time in transferrin.
转铁蛋白是一种参与体液中铁运输的糖蛋白，通过连续阴离子交换色谱法和凝胶过滤从羊水过多患者的羊水中分离得到。人羊水转铁蛋白（hAFT）的N-聚糖经PNGase-F酶切释放，通过凝胶过滤分离，再经（高pH）阴离子交换色谱法分级分离。对天然hAFT进行碱性硼氢化钠处理后，释放的O-聚糖通过凝胶过滤分离，再经阴离子交换色谱法分级分离。通过500或600 MHz 1H-NMR光谱对14种N-聚糖和2种O-聚糖进行结构解析。除了早期为人血清转铁蛋白（hST）确定的常规N-聚糖外，还发现了新的（α1-3）-岩藻糖基化N-聚糖，代表唾液酸化Le（x）元件。此外，与hST相比，检测到更高程度的（α1-6）-岩藻糖基化以及从二天线化合物到三天线化合物分支增加。转铁蛋白中首次证实存在O-聚糖。
