An engineered monomer binding-protein for α-synuclein efficiently inhibits the proliferation of amyloid fibrils
An engineered monomer binding-protein for α-synuclein efficiently inhibits the proliferation of amyloid fibrils
复制标题
DOI：
10.7554/elife.46112
复制
发表时间：
2019-08-21
期刊：
影响因子：
7.7
通讯作者：
Buell, Alexander K.
中科院分区：
文献类型：
--
作者：
Agerschou, Emil Dandanell;Flagmeier, Patrick;Buell, Alexander K.
去除或阻止α-突触核蛋白聚集体的形成是对抗帕金森病的一种可行策略。为此，我们设计了 β-包裹蛋白 AS69，以高亲和力结合单体 α-突触核蛋白。在培养细胞中，AS69 减少突触核蛋白的自身相互作用和可见的 α-突触核蛋白聚集体的形成。在果蝇中，AS69 减少了 α-突触核蛋白聚集以及神经元细胞中 α-突触核蛋白表达导致的运动缺陷。在体外生物物理实验中，即使存在大量过量的单体，AS69 也能高度亚化学计量地抑制初级成核过程和自催化次级成核过程。我们提供的证据表明，AS69-α-突触核蛋白复合物，而不是游离的 AS69，是负责二次成核的亚化学计量抑制的抑制物质。这些结果代表了一种新的范例，即高亲和力单体粘合剂可以导致成核过程的强烈亚化学计量抑制。
Removing or preventing the formation of alpha-synuclein aggregates is a plausible strategy against Parkinson's disease. To this end, we have engineered the beta-wrapin AS69 to bind monomeric alpha-synuclein with high affinity. In cultured cells, AS69 reduced the self-interaction of a synuclein and formation of visible alpha-synuclein aggregates. In flies, AS69 reduced alpha-synuclein aggregates and the locomotor deficit resulting from alpha-synuclein expression in neuronal cells. In biophysical experiments in vitro, AS69 highly sub-stoichiometrically inhibited both primary and autocatalytic secondary nucleation processes, even in the presence of a large excess of monomer. We present evidence that the AS69-alpha-synuclein complex, rather than the free AS69, is the inhibitory species responsible for sub-stoichiometric inhibition of secondary nucleation. These results represent a new paradigm that high affinity monomer binders can lead to strongly sub-stoichiometric inhibition of nucleation processes.