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基于质谱的蛋白质组学方法用于全局和高置信度的蛋白质R-甲基化分析

JoVE Journal
Bioquímica

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JoVE Journal Bioquímica

A Mass Spectrometry-Based Proteomics Approach for Global and High-Confidence Protein R-Methylation Analysis

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基于质谱的蛋白质组学方法用于全局和高置信度的蛋白质R-甲基化分析

DOI:

10.3791/62409-v

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09:40 min

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April 28, 2022

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Marianna Maniaci*^1,2, Federica Marini*¹, Enrico Massignani², Tiziana Bonaldi

¹Department of Experimental Oncology,European Institute of Oncology IRCCS (IEO), ²European School of Molecular Medicine (SEMM), c/o Campus IFOM-IEO

Capítulos

00:04Introduction
00:44Lysate Digestion
02:08Off‐Line HpH‐RP Chromatographic Fractionation of Peptides
02:53R‐Methylated Peptide Immuno‐Affinity Enrichment
06:47Results: R‐Methylation Analysis by Mass Spectrometry
08:39Conclusion

Summary

Tadução automática

蛋白精氨酸（R）-甲基化是一种广泛的翻译后修饰，调节多种生物学途径。质谱法是全球分析R-甲基蛋白质组的最佳技术，当与修饰肽富集的生化方法相结合时。本文介绍了专为高置信度鉴定人细胞中全球R-甲基化而设计的工作流程。

Tags

Keywords: Mass Spectrometry Proteomics Protein R-methylation PRMT Inhibitors DTT Iodoacetamide Trypsin LysargiNase High PH Reversed Phase Liquid Chromatography Immuno-affinity Enrichment

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