de Beer T, van Zuylen C W, Hård K, Boelens R, Kaptein R, Kamerling J P, Vliegenthart J F
Department of Bio-Organic Chemistry, Utrecht University, The Netherlands.
FEBS Lett. 1994 Jul 4;348(1):1-6. doi: 10.1016/0014-5793(94)00547-8.
The structure assessment of an intact glycoprotein in solution requires an extensive assignment of the carbohydrate NMR resonances. However, assignment of homonuclear spectra is very complicated because of the severe overlap of protein and carbohydrate signals. Application of pulsed field gradients allowed high quality natural abundance 1H-13C HSQC and HSQC-TOCSY spectra to be recorded of the alpha-subunit of human chorionic gonadotropin. Most carbohydrate 1H-13C correlations appear in a distinct region between the aromatic region and the protein C alpha-H alpha region. The enormous reduction in overlap led to fast and unambiguous assignment of the anomeric 1H-13C correlations. Subsequently, correlations of the monosaccharide skeleton atoms were readily assigned in the HSQC-TOCSY spectrum.
溶液中完整糖蛋白的结构评估需要对碳水化合物的核磁共振共振进行广泛归属。然而，由于蛋白质和碳水化合物信号的严重重叠，同核谱的归属非常复杂。脉冲场梯度的应用使得能够记录人绒毛膜促性腺激素α亚基的高质量天然丰度1H-13C HSQC和HSQC-TOCSY谱。大多数碳水化合物的1H-13C相关性出现在芳香区和蛋白质Cα-Hα区之间的一个独特区域。重叠的大幅减少使得异头碳的1H-13C相关性能够快速且明确地归属。随后，单糖骨架原子的相关性在HSQC-TOCSY谱中很容易被归属。
