Marks M S, Germain R N, Bonifacino J S
Cell Biology and Metabolism Branch, NICHD, National Institutes of Health, Bethesda, Maryland 20892, USA.
J Biol Chem. 1995 May 5;270(18):10475-81. doi: 10.1074/jbc.270.18.10475.
Many cell surface proteins exist as complexes of multiple subunits. It is well established that most such complexes are assembled within the endoplasmic reticulum (ER). However, the mechanistic details of the assembly process are largely unknown. We show here that alpha and beta subunits of major histocompatibility complex class II antigens in spleen cells of normal mice pass through a transiently aggregated phase in the ER prior to assembly with the invariant chain (Ii). Aggregates form immediately after synthesis and disappear concomitantly with assembly of mature alpha beta Ii complexes. In spleen cells lacking Ii, aggregates fail to be efficiently dissociated over time, implicating subunit assembly as a requirement for disaggregation. Two ER chaperones, BiP and calnexin, bind to newly synthesized class II MHC chains but do not contribute appreciably to the large size of the aggregates. Our observations suggest that some subunits of multisubunit complexes pass through a transient, dynamic high molecular weight aggregate phase during the physiological process of assembly. The results further suggest a novel role for Ii in promoting stable dissociation of preformed aggregates containing alpha and beta subunits rather than in preventing their formation.
许多细胞表面蛋白以多个亚基的复合物形式存在。众所周知,大多数此类复合物是在内质网(ER)中组装的。然而,组装过程的机制细节在很大程度上尚不清楚。我们在此表明,正常小鼠脾细胞中主要组织相容性复合体II类抗原的α和β亚基在与恒定链(Ii)组装之前,在内质网中经历一个短暂的聚集阶段。聚集物在合成后立即形成,并随着成熟的αβIi复合物的组装而同时消失。在缺乏Ii的脾细胞中,随着时间的推移,聚集物不能有效地解离,这表明亚基组装是解离的必要条件。两种内质网伴侣蛋白,BiP和钙连蛋白,与新合成的II类MHC链结合,但对聚集物的大尺寸没有明显贡献。我们的观察结果表明,多亚基复合物的一些亚基在组装的生理过程中会经历一个短暂的、动态的高分子量聚集阶段。结果进一步表明,Ii在促进含有α和β亚基的预先形成的聚集物的稳定解离方面具有新作用,而不是防止它们的形成。