PEPTIDE SYNTHESIS, OLIGOMER PREPARATION AND CONFORMATION-DEPENDENT ANTIBODY CORE

尽管有令人信服的证据表明淀粉样蛋白形成蛋白在疾病中的因果作用，但类型 淀粉样蛋白和其组装状态的关系存在争议，仍有待确定。前纤维淀粉样油- Gomers是直径约3-10 nm的可溶球形聚集体，已观察到 许多不同类型的淀粉样蛋白的电子显微镜和原子力显微镜。这方面的重大进展之一 我们对纤维前低聚物的理解是发现淀粉样寡聚体有一种共同的多肽 一种有别于淀粉样纤维的主干结构，基于构象依赖于- ENT抗体特异性识别淀粉样寡聚体上的共同表位，但不识别纤维、单体或 天然折叠的蛋白质可用于多种不同类型的蛋白质。这表明该抗体识别gE- 独立于氨基酸序列的神经性多肽主干表位，但与 在所有类型的淀粉样寡聚体中很常见。抗低聚物抗体一般也抑制有毒的- 体外检测可溶性低聚物的稳定性。由于不同淀粉样寡聚体具有共同的结构和 它们对细胞通常是有毒的，无论它们来自什么蛋白质，这表明它们 在退行性疾病中具有相同的主要毒性机制。越来越多的证据表明- 研究表明，淀粉样低聚体的膜通透性可能代表了常见的主要机制-- 淀粉样蛋白相关退行性疾病发病机制的NISM 该项目的总体目标是阐明膜通透性的机制，通过 淀粉样蛋白前纤维低聚物。我支持项目的具体目标，核心C将提供同源- 该项目的前纤维低聚物的OU制备以及荧光和氚-AFTOLI- 戈默斯。核心C还将提供替代组装状态的准备，如淀粉样纤维和- 以纤细原纤维为对照。核心C还提供构象依赖的抗体，特异性地识别- 识别用于阻断膜通透性的前纤维低聚物、纤维和环状原纤维 并验证低聚物制剂在实验过程中和实验后的构象状态。

Even though there is compelling evidence for a causal role of amyloid forming proteins in disease, the type of amyloid and its assembly state is controversial and remains to be established. Prefibrillar amyloid oli- gomers are soluble spherical aggregates of approximately 3-10 nm in diameter that have been observed for many different types of amyloids by electron and atomic force microscopy. One of the significant advances in our understanding of prefibrillar oligomers was the finding that amyloid oligomers have a common peptide backbone structure that is distinct from amyloid fibrils based on the observation that a conformation depend- ent antibody specifically recognizes a common epitope on amyloid oligomers, but not fibrils, monomers or natively folded proteins for many different types of proteins. This indicates that the antibody recognizes a ge- neric polypeptide backbone epitope that is independent of the amino acid sequence, but yet is shared in common among all types of amyloid oligomers. The anti-oligomer antibody also generically inhibits the toxic- ity of soluble oligomers examined in vitro. Since different amyloid oligomers share a common structure and they are generically toxic to cells regardless of what protein they are derived from, this suggests that they have the same primary mechanism of toxicity in degenerative diseases. A growing body of evidence sug- gests that membrane permeabilization by amyloid oligomers may represent the common, primary mecha- nism of pathogenesis of amyloid related degenerative diseases. The overall goal of this program project is to elucidate the mechanism of membrane permeabilization by amyloid prefibrillar oligomers. I support of the specific aims of the projects, Core C will provide homogene- ous preparations of prefibrillar Aft oligomers to the projects as well as fluorescent and deuterated Aftoli- gomers. Core C will also provide preparations of alternative assembly states, such as amyloid fibrils and an- nular protofibrils as controls. Core C also provides conformation dependent antibodies that specifically rec- ognize prefibrillar oligomers, fibrils and annular protofibrils for use in blocking membrane permeabilization and verifying the conformational status of the oligomer preparations during and after the experiments.