PURIFICATION AND PARTIAL STRUCTURAL AND FUNCTIONAL-CHARACTERIZATION OF MOUSE MYELIN OLIGODENDROCYTE GLYCOPROTEIN
PURIFICATION AND PARTIAL STRUCTURAL AND FUNCTIONAL-CHARACTERIZATION OF MOUSE MYELIN OLIGODENDROCYTE GLYCOPROTEIN
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DOI：
10.1111/j.1471-4159.1992.tb10040.x
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发表时间：
1992-05-01
影响因子：
4.7
通讯作者：
MATTHIEU, JM
中科院分区：
文献类型：
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作者：
AMIGUET, P;GARDINIER, MV;MATTHIEU, JM
髓鞘/少突胶质细胞糖蛋白（MOG）仅存在于中枢神经系统，定位于髓鞘和少突胶质细胞细胞质膜表面。单克隆抗体8-18C5鉴定MOG。多项研究表明，抗mog抗体可诱导脱髓鞘，从而推断在髓磷脂稳定性中起重要作用。在这项研究中，我们证明了MOG由两个多肽组成，分子质量分别为26和28 kDa。用三氟甲烷磺酸或肽- n4 -（n -乙酰- β -氨基葡萄糖）天冬酰胺酶去糖基化后，这条双线带变成一条25 kda的单条带，表明没有或很少有o键糖，双线带代表不同的糖基化。部分胰蛋白酶的裂解也产生了较低分子量的双峰带，证实了这一观点。用聚丙烯酰胺凝胶电泳纯化MOG，然后电洗脱。得到3个n端序列，包含8 ~ 26个氨基酸。western blot检测发现MOG与小脑可溶性凝集素无结合。MOG似乎不属于信号转导的gtp结合蛋白。在跳跃和颤抖的髓鞘异常突变小鼠中观察到MOG浓度降低，进一步支持其定位于中枢神经系统致密髓鞘。
The myelin/oligodendrocyte glycoprotein (MOG) is found exclusively in the CNS, where it is localized on the surface of myelin and oligodendrocyte cytoplasmic membranes. The monoclonal antibody 8-18C5 identifies MOG. Several studies have shown that anti-MOG antibodies can induce demyelination, thus inferring an important role in myelin stability. In this study, we demonstrate that MOG consists of two polypeptides, with molecular masses of 26 and 28 kDa. This doublet becomes a single 25-kDa band after deglycosylation with trifluoromethanesulfonic acid or peptide-N4-(N-acetyl-beta-glucosaminyl)asparagine amidase, indicating that there are no or few O-linked sugars and that the doublet band represents differential glycosylation. Partial trypsin cleavage, which also gave a doublet band of lower molecular weight, confirmed this idea. MOG was purified by polyacrylamide gel electrophoresis, followed by electroelution. Three N-terminal sequences of eight to 26 amino acids were obtained. By western blot analysis, no binding was found between MOG and cerebellar soluble lectin. MOG does not seem to belong to the signal-transducing GTP-binding proteins. Reduced MOG concentrations were observed in jimpy and quaking dysmyelinating mutant mice, giving further support to its localization in compact myelin of the CNS.