Covalent Modification of Proteins by Osthole Reactive Metabolites using Proteomic Approaches.

IF 2.1 4区医学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Current drug metabolism Pub Date : 2023-01-01 DOI:10.2174/1389200224666230727123006

Yue Zhuo, Huiling Chen, Chenchen Liu, Yida Zhang, Jiansong Fang, Meng Li, Zhendong Wang, Qiyao Jiang, Liangwen Yu, Huafeng Pan, Qi Wang

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Abstract

Background: Osthole (OST) is a bioactive natural coumarin derived from the plant Cnidium monnieri (L.) Cusson fruit (She Chuang Zi), which has various pharmacological and biological activities. OST contains an α,β- unsaturated lactone, which is an electrophilic group that tends to be metabolized into reactive metabolites (RMs). Then, RMs are able to covalently modify nucleophilic amino acid (AA) residues of target proteins. However, few researchers considered the contribution of the covalent modification induced by OST or its metabolites.

Objective: This study aims to investigate the metabolic profile and the metabolites-protein modification of OST.

Methods: The metabolites of OST were qualitatively identified using UHPLC-Q-TOF-MS. The RMs modification patterns and potentially modified AA residues were confirmed by UHPLC-Q-TOF-MS using rat liver microsomes (RLMs) and model AAs. Finally, the modified peptides derived from high-abundance microsomal peptides were separated via nano-LC-Orbitrap-MS, and then RM-modified proteins were identified using a proteome discoverer.

Results: In the presence of RLMs, OST could rapidly be metabolized within 1 h and hardly identified at 4 h. We detected 10 OST metabolites, 13 OST metabolites-NAC (N-acetyl cysteine) adducts, 3 NAL (N-acetyl lysine) adducts, and 11 GSH (glutathione) adducts. Furthermore, 16 RM-modified protein targets were identified, many of which are included in the essential biological processes of OST's anti-Alzheimer's disease (AD) and anti-tumor.

Conclusion: This study provides a novel perspective on the molecular mechanism of OST's pharmacological activities, as well as identifies potential targets for further development and application of OST and other Natural products (NPs).

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利用蛋白质组学方法通过Osthole反应性代谢产物对蛋白质进行共价修饰。

背景：蛇床子是从蛇床子中提取的一种具有生物活性的天然香豆素，具有多种药理和生物活性。OST含有一种α，β-不饱和内酯，这是一种亲电基团，倾向于代谢为反应性代谢产物（RM）。然后，RM能够共价修饰靶蛋白的亲核氨基酸（AA）残基。然而，很少有研究人员考虑OST或其代谢产物诱导的共价修饰的贡献。目的：研究OST的代谢特性及其代谢产物蛋白修饰。方法：采用UHPLC-Q-TOF-MS对OST的代谢产物进行定性鉴定。使用大鼠肝微粒体（RLMs）和模型AA通过UHPLC-Q-TOF-MS证实了RM的修饰模式和潜在的修饰AA残基。最后，通过nano-LC Orbitrap-MS分离来自高丰度微粒体肽的修饰肽，然后使用蛋白质组发现者鉴定RM修饰的蛋白质。结果：在RLMs存在的情况下，OST可以在1小时内快速代谢，而在4小时时几乎无法鉴定。我们检测到10种OST代谢物、13种OST代谢物NAC（N-乙酰半胱氨酸）加合物、3种NAL（N-乙酰赖氨酸）加合产物和11种GSH（谷胱甘肽）加合体。此外，还鉴定了16个RM修饰的蛋白质靶标，其中许多靶标参与了OST抗阿尔茨海默病（AD）和抗肿瘤的基本生物学过程。结论：本研究为OST药理活性的分子机制提供了一个新的视角，并为OST和其他天然产物的进一步开发和应用确定了潜在的靶点。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Current drug metabolism 医学-生化与分子生物学

CiteScore

4.30

自引率

4.30%

发文量

审稿时长

4-8 weeks

期刊介绍： Current Drug Metabolism aims to cover all the latest and outstanding developments in drug metabolism, pharmacokinetics, and drug disposition. The journal serves as an international forum for the publication of full-length/mini review, research articles and guest edited issues in drug metabolism. Current Drug Metabolism is an essential journal for academic, clinical, government and pharmaceutical scientists who wish to be kept informed and up-to-date with the most important developments. The journal covers the following general topic areas: pharmaceutics, pharmacokinetics, toxicology, and most importantly drug metabolism. More specifically, in vitro and in vivo drug metabolism of phase I and phase II enzymes or metabolic pathways; drug-drug interactions and enzyme kinetics; pharmacokinetics, pharmacokinetic-pharmacodynamic modeling, and toxicokinetics; interspecies differences in metabolism or pharmacokinetics, species scaling and extrapolations; drug transporters; target organ toxicity and interindividual variability in drug exposure-response; extrahepatic metabolism; bioactivation, reactive metabolites, and developments for the identification of drug metabolites. Preclinical and clinical reviews describing the drug metabolism and pharmacokinetics of marketed drugs or drug classes.