PROTEIN CHEMICAL AND IMMUNOCYTOCHEMICAL STUDIES OF MENINGOVASCULAR BETA-AMYLOID PROTEIN IN ALZHEIMERS-DISEASE AND NORMAL AGING
PROTEIN CHEMICAL AND IMMUNOCYTOCHEMICAL STUDIES OF MENINGOVASCULAR BETA-AMYLOID PROTEIN IN ALZHEIMERS-DISEASE AND NORMAL AGING
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DOI：
10.1016/0006-8993(88)90673-7
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发表时间：
1988-11-22
期刊：
影响因子：
2.9
通讯作者：
SELKOE, DJ
中科院分区：
文献类型：
--
作者：
JOACHIM, CL;DUFFY, LK;SELKOE, DJ
作为对先前从阿尔茨海默病（AD）脑中纯化的淀粉样斑块核心的分析的比较，我们对从阿尔茨海默病患者脑膜血管中提取的淀粉样纤维进行了蛋白质化学和免疫细胞化学研究。结果进行了比较，从相同制备的馏分老年正常无脑淀粉样血管病或AD的其他显微镜检查结果。对Glenner和Wong的淀粉样蛋白分离方法进行了改进，包括用十二烷基硫酸钠（SDS）提取。从AD脑膜纯化的淀粉样蛋白的凝胶电泳产生以4.2 kDa为中心的条带。对来自AD脑膜的HPLC纯化的淀粉样蛋白的测序证实了已发表的β-残基1-30和35-40的蛋白质序列，除了在位置11的谷氨酸而不是谷氨酰胺。N-末端异质性不显著。没有获得超过残基40的序列。分子量相似但不相同的蛋白质。重量 存在于正常脑膜的HPLC纯化级分中; β-在这些级分中没有检测到蛋白质序列，也没有检测到任何其它可解释的序列。针对纯化的AD脑膜血管淀粉样蛋白提出的两种抗血清在AD制剂的Western印迹上鉴定了4.2 kDa条带;在对照制剂中没有标记该区域的蛋白条带。AD脑膜制备物中的4.2kDa条带也用合成β-内酰胺酶的抗血清标记。肽1 -28的羧基末端的抗血清，而不是通过抗β-肽1 - 28的羧基末端的抗血清。蛋白质前体AD脑膜血管淀粉样蛋白抗血清选择性标记皮质和脑膜血管和斑块核心中的淀粉样蛋白;缠结、斑块神经突和正常CNS细胞以及许多非神经组织未染色。抗血清还标记了血管淀粉样蛋白的偶尔沉积物和在一些无AD的老年个体中发现的不太频繁的斑块核心淀粉样蛋白。我们得出结论：（1）脑膜血管淀粉样蛋白β- AD的蛋白质，其序列已被证实并延伸至残基40，在神经元缠结中不能免疫细胞化学检测到;（2）β-在缺乏光显微镜可见的脑膜血管淀粉样蛋白的老年对照的脑膜制备物中不能检测到蛋白质;和（3）存在于老年正常人中的血管和斑块核心淀粉样蛋白与AD脑膜血管淀粉样蛋白具有抗原交叉反应性。
As a comparison to previous analyses of purified amyloid plaque cores from Alzheimer''s disease (AD) brain, we performed protein chemical and immunocytochemical studies on amyloid filaments extracted from meningeal blood vessels of patients with Alzheimer''s disease. Results were compared with those obtained from identically prepared fractions of aged normals without cerebral amyloid angiopathy or other microscopic findings of AD. The amyloid isolation method of Glenner and Wong was modified, including an extraction with sodium dodecyl sulfate (SDS). Gel electrophoresis of purified amyloid from AD meninges yielded bands centered at 4.2 kDa. Sequencing of the HPLC-purified amyloid protein from AD meninges confirmed the published .beta.-protein sequence for residues 1-30 and 35-40, with the exception of glutamic acid rather than glutamine at position 11. N-terminal heterogeneity was not prominent. No sequence beyond residue 40 was obtained. Proteins of similar but not identical mol. wt. were present in HPLC-purified fractions of normal meninges; neither the .beta.-protein sequence nor any other interpretable sequence was detected in such fractions. The two antisera raised against the purified AD meningovascular amyloid protein identified the 4.2 kDa band on Western blots of AD preparations; no protein band in this region was labeled in control preparations. The 4.2 kDa band in AD meningeal preparations was also labeled by an antiserum to synthetic .beta.-peptide1-28 but not by an antiserum to the carboxyl terminus of the .beta.-protein precursor. Both the AD meningovascular amyloid antisera selectively labeled amyloid in cortical and meningeal vessels and plaque cores; tangles, plaque neurites, and cells of normal CNS and numerous non-neural tissues were unstained . The antisera also labeled the occasional deposits of vascular amyloid and less frequent plaque core amyloid found in some aged individuals without AD. We conclude that (1) the meningovascular amyloid .beta.-protein of AD, whose sequence has been confirmed and extended to residue 40, was not immunocytochemically detectable in neurofibrillary tangles; (2) .beta.-protein could not be detected in meningeal preparations from aged controls who lack light microscopically visible meningovascular amyloid; and (3) the vascular and plaque core amyloid present in aged normals is antigenically cross-reactive with AD meningovascular amyloid.