Immunochemical identification of ubiquitin and heat-shock proteins in corpora amylacea from normal aged and Alzheimer's disease brains.
Immunochemical identification of ubiquitin and heat-shock proteins in corpora amylacea from normal aged and Alzheimer's disease brains.
复制标题
正常老年人和阿尔茨海默病大脑淀粉体中泛素和热休克蛋白的免疫化学鉴定。
DOI：
10.1007/bf00227716
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发表时间：
1993
影响因子：
12.7
通讯作者：
Gauvreau,D
中科院分区：
文献类型：
--
作者：
Cissé,S;Perry,G;Lacoste-Royal,G;Cabana,T;Gauvreau,D
淀粉体(CA)在中枢神经系统(CNS)中的积聚与正常衰老和神经退行性疾病(如阿尔茨海默病(AD))有关。据报道，CA主要由葡萄糖聚合物组成，但CA总重量的约4%始终由蛋白质组成。CA蛋白经十二烷基硫酸钠-聚丙烯酰胺凝胶电泳法分离后，显示24~133 kDa的多肽，有4条丰富的条带。从纯化的CA中溶解的多肽的免疫印迹显示，所有条带都呈泛素(Ub)阳性。热休克蛋白(HSP)28和70的抗血清选择性地与30和67 kDa的条带反应。这些结果表明，Ub与CA的主要蛋白质组分有关，多肽可能是Ub的偶联物。免疫组织化学染色实验对脑组织切片中CA的蛋白质组分以及从AD和正常老年脑中提纯的CA的蛋白质组分进行了特异性的表征。在所有病例中，CA与Ub抗体呈阳性反应，与成对螺旋细丝或HSP28或70的抗体呈阳性反应，其中最显著的染色是Ub、HSP28或HSP70的抗体。Ub和HSP28和70在应激后被主动诱导，这表明改变的蛋白质的积累可能归因于异常翻译后修饰的频率增加或持续的生理应激(与正常衰老和神经退行性过程有关)，可能参与了CA的发病。
Corpora amylacea (CA) accumulation in the central nervous system (CNS) is associated with both normal aging and neurodegenerative conditions such as Alzheimer's disease (AD). CA is reported to be primarily composed of glucose polymers, but approximately 4% of the total weight of CA is consistently composed of protein. CA protein resolved on sodium dodecylsulfatepolyacrylamide gel electrophoresis showed a broad range of polypeptides ranging from 24 to 133 kDa, with four abundant bands. Immunoblots of the profile of polypeptides solubilized from purified CA, showed positive ubiquitin (Ub) immunoreactivity for all the bands. Antisera to heat-shock proteins (hsp) 28 and 70 reacted selectively with bands of 30 and 67 kDa. These results show that Ub is associated with the primary protein components of CA and that the polypeptides are likely to be Ub conjugates. Immunostaining experiments were performed to specifically characterize the protein components of CA in brain tissue sections as well as those of CA purified from both AD and normal aged brains. In all cases CA showed positive reactions with antibodies to Ub, with antibodies raised against either paired helical filaments or hsp 28 or 70, the most prominent staining being with antibodies to Ub, hsp 28 or hsp 70. The presence of Ub and hsp 28 and 70, which are actively induced after stress, suggests that accumulation of altered proteins, possibly attributed to an increased frequency of unusual post-translational modifications or to a sustained physiological stress (related to both normal aging and neurodegenerative process), may be involved in the pathogenesis of CA.