Bonnon Carine, Bel Christophe, Goutebroze Laurence, Maigret Bernard, Girault Jean-Antoine, Faivre-Sarrailh Catherine
Neurobiologie des Interactions Cellulaires et Neurophysiopathologie, CNRS UMR 6184, Université de la Méditerranée, Institut Jean-Roche, 13916 Marseille Cedex 20, France.
Mol Biol Cell. 2007 Jan;18(1):229-41. doi: 10.1091/mbc.e06-06-0570. Epub 2006 Nov 8.
Formation of nodes of Ranvier requires contact of axons with myelinating glial cells, generating specialized axo-glial subdomains. Caspr/paranodin is required for the formation of septate-like junctions at paranodes, whereas the related caspr2 is essential for the organization of juxtaparanodes. The molecular mechanisms underlying the segregation of these related glycoproteins within distinct complexes are poorly understood. Exit of paranodin from the endoplasmic reticulum (ER) is mediated by its interaction with F3/contactin. Using domain swapping with caspr2, we mapped a motif with Pro-Gly-Tyr repeats (PGY) in the ectodomain of paranodin responsible for its ER retention. Deletion of PGY allows cell surface delivery of paranodin bypassing the calnexin-calreticulin quality control. Conversely, insertion of PGY in caspr2 or NrCAM blocks these proteins in the ER. PGY is a novel type of processing signal that compels chaperoning of paranodin by contactin. Contactin associated with paranodin is expressed at the cell surface with high-mannose N-glycans. Using mutant CHO lines altered in the processing of N-linked carbohydrates, we show that the high-mannose glycoform of contactin strongly binds neurofascin-155, its glial partner at paranodes. Thus, the unconventional processing of paranodin and contactin may determine the selective association of axo-glial complexes at paranodes.
郎飞结的形成需要轴突与形成髓鞘的神经胶质细胞接触,从而产生特化的轴突-神经胶质亚结构域。Caspr/ paranodin是在 paranodes处形成类紧密连接所必需的,而相关的caspr2对于旁结的组织至关重要。这些相关糖蛋白在不同复合物中分离的分子机制尚不清楚。paranodin从内质网(ER)的输出是由其与F3/contactin的相互作用介导的。通过与caspr2进行结构域交换,我们在paranodin的胞外结构域中定位了一个具有Pro-Gly-Tyr重复序列(PGY)的基序,该基序负责其在内质网中的保留。删除PGY可使paranodin绕过钙连蛋白-钙网蛋白质量控制而在细胞表面递送。相反,在caspr2或NrCAM中插入PGY会将这些蛋白质滞留在内质网中。PGY是一种新型的加工信号,它迫使contactin对paranodin进行伴侣作用。与paranodin相关的contactin在细胞表面以高甘露糖N-聚糖形式表达。使用在N-连接碳水化合物加工过程中发生改变的突变CHO细胞系,我们发现contactin的高甘露糖糖型强烈结合神经束蛋白-155,后者是其在paranodes处的神经胶质伴侣。因此,paranodin和contactin的非常规加工可能决定了paranodes处轴突-神经胶质复合物的选择性结合。