Suzuki A, Sugiyama Y, Hayashi Y, Nyu-i N, Yoshida M, Nonaka I, Ishiura S, Arahata K, Ohno S
Department of Molecular Biology, Yokohama City University School of Medicine, Kanazawa-ku, Yokohama 236, Japan.
J Cell Biol. 1998 Mar 9;140(5):1113-24. doi: 10.1083/jcb.140.5.1113.
Muscle cells are frequently subjected to severe conditions caused by heat, oxidative, and mechanical stresses. The small heat shock proteins (sHSPs) such as alphaB-crystallin and HSP27, which are highly expressed in muscle cells, have been suggested to play roles in maintaining myofibrillar integrity against such stresses. Here, we identified a novel member of the sHSP family that associates specifically with myotonic dystrophy protein kinase (DMPK). This DMPK-binding protein, MKBP, shows a unique nature compared with other known sHSPs: (a) In muscle cytosol, MKBP exists as an oligomeric complex separate from the complex formed by alphaB-crystallin and HSP27. (b) The expression of MKBP is not induced by heat shock, although it shows the characteristic early response of redistribution to the insoluble fraction like other sHSPs. Immunohistochemical analysis of skeletal muscle cells shows that MKBP localizes to the cross sections of individual myofibrils at the Z-membrane as well as the neuromuscular junction, where DMPK has been suggested to be concentrated. In vitro, MKBP enhances the kinase activity of DMPK and protects it from heat-induced inactivation. These results suggest that MKBP constitutes a novel stress-responsive system independent of other known sHSPs in muscle cells and that DMPK may be involved in this system by being activated by MKBP. Importantly, since the amount of MKBP protein, but not that of other sHSP family member proteins, is selectively upregulated in skeletal muscle from DM patients, an interaction between DMPK and MKBP may be involved in the pathogenesis of DM.
肌肉细胞经常受到由热、氧化和机械应力引起的严峻条件影响。诸如αB-晶状体蛋白和HSP27等小热休克蛋白(sHSPs)在肌肉细胞中高度表达,有人认为它们在维持肌原纤维完整性以抵抗此类应力方面发挥作用。在此,我们鉴定出sHSP家族的一个新成员,它与强直性肌营养不良蛋白激酶(DMPK)特异性结合。这种与DMPK结合的蛋白,即MKBP,与其他已知的sHSPs相比具有独特的性质:(a)在肌肉胞质溶胶中,MKBP以寡聚复合物的形式存在,与由αB-晶状体蛋白和HSP27形成的复合物分开。(b)MKBP的表达不受热休克诱导,尽管它像其他sHSPs一样表现出重新分布到不溶性部分的特征性早期反应。骨骼肌细胞的免疫组织化学分析表明,MKBP定位于单个肌原纤维在Z膜处的横截面以及神经肌肉接头处,而DMPK被认为在这些部位聚集。在体外,MKBP增强DMPK的激酶活性并保护其免受热诱导的失活。这些结果表明,MKBP在肌肉细胞中构成了一个独立于其他已知sHSPs的新的应激反应系统,并且DMPK可能通过被MKBP激活而参与该系统。重要的是,由于DM患者骨骼肌中MKBP蛋白的量而非其他sHSP家族成员蛋白的量被选择性上调,DMPK与MKBP之间的相互作用可能参与了DM的发病机制。