Half-Time Heat Map Reveals Ultrasonic Effects on Morphology and Kinetics of Amyloidogenic Aggregation Reaction
Half-Time Heat Map Reveals Ultrasonic Effects on Morphology and Kinetics of Amyloidogenic Aggregation Reaction
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半时热图揭示超声波对淀粉样蛋白聚集反应的形态和动力学的影响
DOI：
10.1021/acschemneuro.1c00461
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发表时间：
2021
影响因子：
5
通讯作者：
Ogi Hirotsugu
中科院分区：
文献类型：
--
作者：
Nakajima Kichitaro;Toda Hajime;Yamaguchi Keiichi;So Masatomo;Ikenaka Kensuke;Mochizuki Hideki;Goto Yuji;Ogi Hirotsugu
超声波能加速淀粉样纤维的形成，在淀粉样变性的早期诊断中有着广阔的应用前景。虽然该技术在蛋白质科学领域的应用正在扩大，但其对淀粉样蛋白聚集反应的影响知之甚少。在这项研究中，我们全面研究了所得的聚集体的形态和结构，原纤维形成的动力学，以及在超声处理下使用β2-微球蛋白的种子检测灵敏度，并在振荡下比较了这些特性，这在淀粉样蛋白原纤维测定中传统上被采用。为了讨论超声波对淀粉样纤维形成的影响，我们提出了半时热图，它描述了淀粉样蛋白聚集反应的相图。实验结果表明，超声波处理大大促进原纤维的形成，特别是在稀单体溶液中，诱导短分散的原纤维，并能够检测超痕量浓度的种子，检测限为10 fM。此外，我们指出，超声波处理高度改变的能量景观的聚集反应，由于超声波空化的影响。这些见解不仅有助于我们理解搅拌对淀粉样蛋白聚集反应的影响，而且有助于其在淀粉样蛋白病临床诊断中的有效应用。
Ultrasonication has been recently adopted in amyloid-fibril assays because of its ability to accelerate fibril formation, being promising in the early stage diagnosis of amyloidoses in clinical applications. Although applications of this technique are expanding in the field of protein science, its effects on the aggregation reactions of amyloidogenic proteins are poorly understood. In this study, we comprehensively investigated the morphology and structure of resultant aggregates, kinetics of fibril formation, and seed-detection sensitivity under ultrasonication using β2-microglobulin and compared these characteristics under shaking, which has been traditionally adopted in amyloid-fibril assays. To discuss the ultrasonic effects on the amyloid-fibril formation, we propose the half-time heat map, which describes the phase diagram of the aggregation reaction of amyloidogenic proteins. The experimental results show that ultrasonication greatly promotes fibril formation, especially in dilute monomer solutions, induces short-dispersed fibrils, and is capable of detecting ultra-trace-concentration seeds with a detection limit of 10 fM. Furthermore, we indicate that ultrasonication highly alters the energy landscape of an aggregation reaction due to the effect of ultrasonic cavitation. These insights contribute not only to our understanding of the effects of agitation on amyloidogenic aggregation reactions but also to their effective application in the clinical diagnosis of amyloidoses.