Shimizu H, Ochiai K, Ikenaka K, Mikoshiba K, Hase S
Department of Chemistry, Osaka University College of Science.
J Biochem. 1993 Sep;114(3):334-8. doi: 10.1093/oxfordjournals.jbchem.a124177.
Sugar chains were liberated from mouse tissues (liver, heart, spleen, kidney, thymus, cerebrum, cerebellum, and brain stem) by hydrazinolysis. After acetylation of the free amino groups, the sugar chains released were pyridylaminated. Pyridylamino (PA-) derivatives of the sugar chains from each tissue were fractionated by anion-exchange high-performance liquid chromatography (HPLC) according to their negative charges. Fractions containing PA-neutral sugar chains thus obtained were separated successively by size-fractionation HPLC and reversed-phase HPLC. Two sugar chains that were more abundant in neural tissues than in other tissues were purified. The structures of the two sugar chains were determined by sugar composition analysis, sequential exoglycosidase digestion, and methylation analysis. The proposed structures are shown below. A structure with n = 1 was found in the cerebrum, cerebellum, and brain stem. A structure with n = 0 was abundant in the cerebrum and brain stem, but less so in the cerebellum. [formula: see text]
通过肼解从小鼠组织(肝脏、心脏、脾脏、肾脏、胸腺、大脑、小脑和脑干)中释放糖链。游离氨基乙酰化后,将释放的糖链进行吡啶基胺化。来自每个组织的糖链的吡啶基氨基(PA-)衍生物根据其负电荷通过阴离子交换高效液相色谱(HPLC)进行分级分离。由此获得的含有PA-中性糖链的级分依次通过尺寸分级HPLC和反相HPLC进行分离。纯化了两种在神经组织中比在其他组织中更丰富的糖链。通过糖组成分析、外切糖苷酶顺序消化和甲基化分析确定了这两种糖链的结构。所提出的结构如下所示。在大脑、小脑和脑干中发现了n = 1的结构。n = 0的结构在大脑和脑干中丰富,但在小脑中较少。[分子式:见正文]