劈理(地质)
化学
分子动力学
细胞生物学
生物物理学
基质(水族馆)
底物特异性
动力学(音乐)
淀粉样前体蛋白分泌酶
神经科学
立体化学
生物
生物化学
淀粉样前体蛋白
心理学
酶
医学
内科学
计算化学
阿尔茨海默病
古生物学
疾病
断裂(地质)
生态学
教育学
作者
N. Hung,Shweta R. Malvankar,Michael S. Wolfe,Yinglong Miao
标识
DOI:10.1021/acschemneuro.3c00594
摘要
γ-Secretase is an intramembrane aspartyl protease complex that cleaves the transmembrane domain of over 150 peptide substrates, including amyloid precursor protein (APP) and the Notch family of receptors, via two conserved aspartates D257 and D385 in the presenilin-1 (PS1) catalytic subunit. However, while the activation of γ-secretase for cleavage of APP has been widely studied, the cleavage of Notch by γ-secretase remains poorly explored. Here, we combined Gaussian accelerated molecular dynamics (GaMD) simulations and mass spectrometry (MS) analysis of proteolytic products to present the first dynamic models for cleavage of Notch by γ-secretase. MS showed that γ-secretase cleaved the WT Notch at Notch residue G34, while cleavage of the L36F mutant Notch occurred at Notch residue C33. Initially, we prepared our simulation systems starting from the cryoEM structure of Notch-bound γ-secretase (PDB: 6IDF) and failed to capture the proper cleavages of WT and L36F Notch by γ-secretase. We then discovered an incorrect registry of the Notch substrate in the PS1 active site through alignment of the experimental structure of Notch-bound (PDB: 6IDF) and APP-bound γ-secretase (PDB: 6IYC). Every residue of the APP substrate was systematically mutated to the corresponding Notch residue to prepare a resolved model of Notch-bound γ-secretase complexes. GaMD simulations of the resolved model successfully captured γ-secretase activation for proper cleavages of both WT and L36F mutant Notch. Our findings presented here provided mechanistic insights into the structural dynamics and enzyme–substrate interactions required for γ-secretase activation for cleavage of Notch and other substrates.
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