In vivo aggregation of beta-amyloid peptide variants.
In vivo aggregation of beta-amyloid peptide variants.
复制标题
β-淀粉样肽变体的体内聚集。
DOI：
10.1046/j.1471-4159.1998.71041616.x
复制
发表时间：
1998
影响因子：
4.7
通讯作者：
Link,CD
中科院分区：
文献类型：
--
作者：
Fay,DS;Fluet,A;Johnson,CJ;Link,CD
转基因秀丽隐杆线虫动物经过工程改造，可表达长形式的人类 β-淀粉样肽 (Aβ 1-42) 的野生型和单氨基酸变体。通过定量免疫印迹测定，这些动物表达高水平的全长肽（约 300 ng Aβ/mg 总蛋白）。这些动物中野生型 Aβ 的表达导致与刚果红和硫代黄素 S 反应的淀粉样沉积物的快速产生。该模型系统已用于检查 Leu17Pro、Leu17Val、Ala30-Pro、Met35Cys 和 Met35Leu 替代对体内淀粉样沉积物产生的影响。我们发现 Leu17Pro 和 Met35Cys 取代完全阻止了硫黄素 S 反应沉积物的形成，表明它们是体内淀粉样蛋白形成的关键残基。我们还构建了表达新型 Aβ 变体（单链二聚体）的转基因菌株。表达高水平这种变体的动物也无法产生硫代黄素 S 反应性沉积物。
TransgenicCaenorhabditis elegansanimals have been engineered to express wild‐type and single‐amino acid variants of a long form of human β‐amyloid peptide (Aβ 1–42). These animals express high levels (∼300 ng of Aβ/mg of total protein) of apparently full‐length peptide, as determined by quantitative immunoblot. Expression of wild‐type Aβ in these animals leads to rapid production of amyloid deposits reactive with Congo red and thioflavin S. This model system has been used to examine the effect of Leu17Pro, Leu17Val, Ala30‐Pro, Met35Cys, and Met35Leu substitutions on the in vivo production of amyloid deposits. We find that the Leu17Pro and Met35Cys substitutions completely block the formation of thioflavin S‐reactive deposits, implicating these as key residues for in vivo amyloid formation. We have also constructed transgenic strains expressing a novel Aβ variant, the single‐chain dimer. Animals expressing high levels of this variant also fail to produce thioflavin S‐reactive deposits.