Yamaguchi Y, Takizawa T, Kato K, Arata Y, Shimada I
Graduate School of Pharmaceutical Sciences, The University of Tokyo, Japan.
J Biomol NMR. 2000 Dec;18(4):357-60. doi: 10.1023/a:1026776721348.
In order to understand the role of the glycans in glycoproteins in solution, structural information obtained by NMR spectroscopy is obviously required. However, the assignment of the NMR signals from the glycans in larger glycoproteins is still difficult, mainly due to the lack of appropriate methods for the assignment of the resonances originating from the glycans. By using [U-13C6, 2H7]glucose as a metabolic precursor, we have successfully prepared a glycoprotein whose glycan is uniformly labeled with 13C and partially with D at the sugar residues. The D to H exchange ratios at the C1-C6 positions of the sugar residues have been proven to provide useful information for the spectral assignments of the glycan in the glycoprotein. This is the first report on the residue-specific assignment of the anomeric resonances originating from a glycan attached to a glycoprotein by using the metabolic incorporation of hydrogen from the medium into a glycan labeled with [U-13C6, 2H7]glucose.
为了了解溶液中糖蛋白聚糖的作用，显然需要通过核磁共振波谱法获得结构信息。然而，在较大的糖蛋白中，聚糖的核磁共振信号归属仍然困难，主要是由于缺乏合适的方法来归属源自聚糖的共振信号。通过使用[U-13C6, 2H7]葡萄糖作为代谢前体，我们成功制备了一种糖蛋白，其聚糖在糖残基上被13C均匀标记且部分被D标记。已证明糖残基C1-C6位置的D与H交换比率可为糖蛋白中聚糖的光谱归属提供有用信息。这是首次通过将培养基中的氢代谢掺入用[U-13C6, 2H7]葡萄糖标记的聚糖中，对源自与糖蛋白相连的聚糖的异头物共振进行残基特异性归属的报告。
