Institute of Bioengineering and Nanotechnology, Singapore 138669.
Proc Natl Acad Sci U S A. 2013 Jan 8;110(2):519-24. doi: 10.1073/pnas.1217742110. Epub 2012 Dec 24.
The self-assembly of abnormally folded proteins into amyloid fibrils is a hallmark of many debilitating diseases, from Alzheimer's and Parkinson diseases to prion-related disorders and diabetes type II. However, the fundamental mechanism of amyloid aggregation remains poorly understood. Core sequences of four to seven amino acids within natural amyloid proteins that form toxic fibrils have been used to study amyloidogenesis. We recently reported a class of systematically designed ultrasmall peptides that self-assemble in water into cross-β-type fibers. Here we compare the self-assembly of these peptides with natural core sequences. These include core segments from Alzheimer's amyloid-β, human amylin, and calcitonin. We analyzed the self-assembly process using circular dichroism, electron microscopy, X-ray diffraction, rheology, and molecular dynamics simulations. We found that the designed aliphatic peptides exhibited a similar self-assembly mechanism to several natural sequences, with formation of α-helical intermediates being a common feature. Interestingly, the self-assembly of a second core sequence from amyloid-β, containing the diphenylalanine motif, was distinctly different from all other examined sequences. The diphenylalanine-containing sequence formed β-sheet aggregates without going through the α-helical intermediate step, giving a unique fiber-diffraction pattern and simulation structure. Based on these results, we propose a simplified aliphatic model system to study amyloidosis. Our results provide vital insight into the nature of early intermediates formed and suggest that aromatic interactions are not as important in amyloid formation as previously postulated. This information is necessary for developing therapeutic drugs that inhibit and control amyloid formation.
异常折叠的蛋白质自组装成淀粉样纤维是许多衰弱性疾病的标志,包括阿尔茨海默病、帕金森病、朊病毒相关疾病和 2 型糖尿病等。然而,淀粉样聚集的基本机制仍知之甚少。天然淀粉样蛋白中形成毒性纤维的四到七个氨基酸的核心序列已被用于研究淀粉样蛋白形成。我们最近报道了一类系统设计的超小肽,它们在水中自组装成交叉-β型纤维。在这里,我们将这些肽与天然核心序列进行比较。其中包括来自阿尔茨海默病淀粉样β、人胰岛淀粉样多肽和降钙素的核心片段。我们使用圆二色性、电子显微镜、X 射线衍射、流变学和分子动力学模拟来分析自组装过程。我们发现,设计的脂肪肽表现出与几种天然序列相似的自组装机制,形成α-螺旋中间体是一个共同特征。有趣的是,来自淀粉样β的第二个核心序列(含有二苯丙氨酸基序)的自组装与所有其他检查的序列明显不同。含有二苯丙氨酸的序列没有经过α-螺旋中间体步骤就形成β-折叠聚集体,给出了独特的纤维衍射图案和模拟结构。基于这些结果,我们提出了一个简化的脂肪模型系统来研究淀粉样变性。我们的结果提供了对形成的早期中间体本质的重要见解,并表明芳香族相互作用在淀粉样形成中并不像以前假设的那样重要。这些信息对于开发抑制和控制淀粉样形成的治疗药物是必要的。