Institut für Physikalische Biologie, Heinrich Heine University Düsseldorf, Düsseldorf, Germany.
Department of Chemistry, University of Cambridge, Cambridge, United Kingdom.
Elife. 2019 Aug 21;8:e46112. doi: 10.7554/eLife.46112.
Removing or preventing the formation of [Formula: see text]-synuclein aggregates is a plausible strategy against Parkinson's disease. To this end, we have engineered the [Formula: see text]-wrapin AS69 to bind monomeric [Formula: see text]-synuclein with high affinity. In cultured cells, AS69 reduced the self-interaction of [Formula: see text]-synuclein and formation of visible [Formula: see text]-synuclein aggregates. In flies, AS69 reduced [Formula: see text]-synuclein aggregates and the locomotor deficit resulting from [Formula: see text]-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-[Formula: see text]-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.
去除或阻止[公式：见文本]-突触核蛋白聚集体的形成是对抗帕金森病的一种合理策略。为此，我们设计了[公式：见文本]-包裹的 AS69 以高亲和力结合单体[公式：见文本]-突触核蛋白。在培养的细胞中，AS69 减少了[公式：见文本]-突触核蛋白的自我相互作用和可见[公式：见文本]-突触核蛋白聚集体的形成。在果蝇中，AS69 减少了[公式：见文本]-突触核蛋白聚集体和神经元细胞中[公式：见文本]-突触核蛋白表达导致的运动缺陷。在体外的生物物理实验中，AS69 以亚化学计量的方式高度抑制了一级和自动催化的二级成核过程，即使在存在大量单体的情况下也是如此。我们提供的证据表明，AS69-[公式：见文本]-突触核蛋白复合物而不是游离的 AS69 是负责亚化学计量抑制二级成核的抑制性物质。这些结果代表了一个新的范例，即高亲和力单体结合物可以导致成核过程的强烈亚化学计量抑制。
