适用于肌肉组织蛋白质组分析的蛋白质提取方法。

IF 4 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY Proteomes Pub Date : 2024-09-25 DOI:10.3390/proteomes12040027
Lorenza Vantaggiato, Claudia Landi, Enxhi Shaba, Daniela Rossi, Vincenzo Sorrentino, Luca Bini
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引用次数: 0

摘要

肌肉组织是哺乳动物体内最具活力和可塑性的组织之一,具有不同的作用,如产生力量和控制新陈代谢。肌肉蛋白质组学为揭示肌肉病理生理学背后的分子机制提供了重要机会。为确保蛋白质组学分析的成功,有必要采用高效且可重复的蛋白质提取方法。本研究旨在评估两种不同的肌肉样本提取方案在二维凝胶电泳中的功效。其中,小鼠肌肉蛋白质的提取分别采用基于 SDS 的缓冲液(方法 A)和UREA/CHAPS/DTE/TRIS 溶液(方法 B)。通过对 2DE 凝胶进行图像分析以及统计和多元分析,评估了这两种方法的有效性。两种制备方法的 2DE 凝胶都显示出良好的分辨率和良好的斑点重叠。图像分析表明两种方法提取的蛋白质丰度不同。结果表明,这两种方法提取和溶解的蛋白质具有不同的化学物理特性和不同的细胞定位。这些结果证明了两种蛋白质提取方法的有效性和可重复性,可同时用于肌肉组织的全面蛋白质组分析。
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Protein Extraction Methods Suitable for Muscle Tissue Proteomic Analysis.

Muscle tissue is one of the most dynamic and plastic tissues of the mammalian body and covers different roles, such as force generation and metabolic control. Muscular proteomics provides an important opportunity to reveal the molecular mechanisms behind muscle pathophysiology. To ensure successful proteomic analysis, it is necessary to have an efficient and reproducible protein extraction method. This study aimed to evaluate the efficacy of two different extraction protocols of muscle samples for two-dimensional gel electrophoresis. In particular, mouse muscle proteins were extracted by an SDS-based buffer (Method A) and by a UREA/CHAPS/DTE/TRIS solution (Method B). The efficacies of the methods were assessed by performing an image analysis of the 2DE gels and by statistical and multivariate analyses. The 2DE gels in both preparations showed good resolution and good spot overlapping. Methods A and B produced 2DE gels with different means of total spots, higher for B. Image analysis showed different patterns of protein abundance between the protocols. The results showed that the two methods extract and solubilize proteins with different chemical-physical characteristics and different cellular localizations. These results attest the efficacy and reproducibility of both protein extraction methods, which can be parallelly applied for comprehensive proteomic profiling of muscle tissue.

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来源期刊
Proteomes
Proteomes Biochemistry, Genetics and Molecular Biology-Clinical Biochemistry
CiteScore
6.50
自引率
3.00%
发文量
37
审稿时长
11 weeks
期刊介绍: Proteomes (ISSN 2227-7382) is an open access, peer reviewed journal on all aspects of proteome science. Proteomes covers the multi-disciplinary topics of structural and functional biology, protein chemistry, cell biology, methodology used for protein analysis, including mass spectrometry, protein arrays, bioinformatics, HTS assays, etc. Our aim is to encourage scientists to publish their experimental and theoretical results in as much detail as possible. Therefore, there is no restriction on the length of papers. Scope: -whole proteome analysis of any organism -disease/pharmaceutical studies -comparative proteomics -protein-ligand/protein interactions -structure/functional proteomics -gene expression -methodology -bioinformatics -applications of proteomics
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