FrozONE:快速富集细胞核,对冷冻组织进行全面的蛋白质组学分析。

IF 3.3 2区 生物学 Q1 BIOLOGY
Life Science Alliance Pub Date : 2024-12-12 Print Date: 2025-03-01 DOI:10.26508/lsa.202403130
Lukas A Huschet, Fabian P Kliem, Peter Wienand, Claudia M Wunderlich, Andrea Ribeiro, Isabel Bustos-Martínez, Ángel Barco, F Thomas Wunderlich, Maciej Lech, Maria S Robles
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引用次数: 0

摘要

亚细胞分馏可用于研究单个细胞器的分区过程。核富集方法通常使用密度梯度结合超速离心来处理新鲜分离的组织。虽然这种方法与蛋白质组学结合使用的范围很广,但在可扩展性和冷冻材料的适用性方面存在一些挑战。为了克服这些限制,我们开发了 FrozONE(冷冻器官细胞核富集),这是一种用于冷冻组织的细胞核富集和蛋白质组学工作流程。通过将我们的工作流程与其他方法进行广泛比对,我们发现 FrozONE 是一种比传统超速离心法更快、更简单、更可扩展的替代方法。FrozONE 可以对具有复杂细胞异质性的不同组织的核蛋白质组进行研究、分析和分类,确保从不同类型的细胞中获得最佳的细胞核富集,并对低含量蛋白质进行定量分析。除了在健康小鼠组织中的表现外,FrozONE 在病理条件下(饮食诱导的非酒精性脂肪性肝炎)的肝核蛋白质组变化特征描述中也被证明是非常有效的。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
FrozONE: quick cell nucleus enrichment for comprehensive proteomics analysis of frozen tissues.

Subcellular fractionation allows for the investigation of compartmentalized processes in individual cellular organelles. Nuclear enrichment methods commonly employ the use of density gradients combined with ultracentrifugation for freshly isolated tissues. Although it is broadly used in combination with proteomics, this approach poses several challenges when it comes to scalability and applicability for frozen material. To overcome these limitations, we developed FrozONE (Frozen Organ Nucleus Enrichment), a nucleus enrichment and proteomics workflow for frozen tissues. By extensively benchmarking our workflow against alternative methods, we showed that FrozONE is a faster, simpler, and more scalable alternative to conventional ultracentrifugation methods. FrozONE allowed for the study, profiling, and classification of nuclear proteomes in different tissues with complex cellular heterogeneity, ensuring optimal nucleus enrichment from different cell types and quantitative resolution for low abundant proteins. In addition to its performance in healthy mouse tissues, FrozONE proved to be very efficient for the characterization of liver nuclear proteome alterations in a pathological condition, diet-induced nonalcoholic steatohepatitis.

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来源期刊
Life Science Alliance
Life Science Alliance Agricultural and Biological Sciences-Plant Science
CiteScore
5.80
自引率
2.30%
发文量
241
审稿时长
10 weeks
期刊介绍: Life Science Alliance is a global, open-access, editorially independent, and peer-reviewed journal launched by an alliance of EMBO Press, Rockefeller University Press, and Cold Spring Harbor Laboratory Press. Life Science Alliance is committed to rapid, fair, and transparent publication of valuable research from across all areas in the life sciences.
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