化学
氢-氘交换
单体
毫秒
共核细胞病
动力学
质谱法
纤维
分子动力学
生物物理学
结晶学
α-突触核蛋白
计算化学
生物化学
有机化学
色谱法
医学
物理
疾病
病理
天文
量子力学
帕金森病
生物
聚合物
作者
Neeleema Seetaloo,Maria Zacharopoulou,Amberley D. Stephens,Gabriele S Kaminski Schierle,Jonathan J. Phillips
出处
期刊:Analytical Chemistry
[American Chemical Society]
日期:2022-11-22
卷期号:94 (48): 16711-16719
被引量:10
标识
DOI:10.1021/acs.analchem.2c03183
摘要
In Parkinson's disease and other synucleinopathies, α-synuclein misfolds and aggregates. Its intrinsically disordered nature, however, causes it to adopt several meta-stable conformations stabilized by internal hydrogen bonding. Because they interconvert on short timescales, monomeric conformations of disordered proteins are difficult to characterize using common structural techniques. Few techniques can measure the conformations of monomeric α-synuclein, including millisecond hydrogen/deuterium-exchange mass spectrometry (HDX-MS). Here, we demonstrate a new approach correlating millisecond HDX-MS data with aggregation kinetics to determine the localized structural dynamics that underpin the self-assembly process in full-length wild-type monomeric α-synuclein. Our custom instrumentation and software enabled measurement of the amide hydrogen-exchange rates on the millisecond timescale for wild-type α-synuclein monomer up to residue resolution and under physiological conditions, mimicking those in the extracellular, intracellular, and lysosomal cellular compartments. We applied an empirical correction to normalize measured hydrogen-exchange rates and thus allow comparison between drastically different solution conditions. We characterized the aggregation kinetics and morphology of the resulting fibrils and correlate these with structural changes in the monomer. Applying a correlative approach to connect molecular conformation to aggregation in α-synuclein for the first time, we found that the central C-terminal residues of α-synuclein are driving its nucleation and thus its aggregation. We provide a new approach to link the local structural dynamics of intrinsically disordered proteins to functional attributes, which we evidence with new details on our current understanding of the relationship between the local chemical environment and conformational ensemble bias of monomeric α-synuclein.
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