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...
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MDPI AG
2024-09-01
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| Series: | Proteomes |
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| Online Access: | https://www.mdpi.com/2227-7382/12/4/27 |
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| author | Lorenza Vantaggiato Claudia Landi Enxhi Shaba Daniela Rossi Vincenzo Sorrentino Luca Bini |
| author_facet | Lorenza Vantaggiato Claudia Landi Enxhi Shaba Daniela Rossi Vincenzo Sorrentino Luca Bini |
| author_sort | Lorenza Vantaggiato |
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| description | 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. |
| format | Article |
| id | doaj-art-c2587099f4a644e89e8ef4ba0dd97398 |
| institution | OA Journals |
| issn | 2227-7382 |
| language | English |
| publishDate | 2024-09-01 |
| publisher | MDPI AG |
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| series | Proteomes |
| spelling | doaj-art-c2587099f4a644e89e8ef4ba0dd973982025-08-20T02:01:14ZengMDPI AGProteomes2227-73822024-09-011242710.3390/proteomes12040027Protein Extraction Methods Suitable for Muscle Tissue Proteomic AnalysisLorenza Vantaggiato0Claudia Landi1Enxhi Shaba2Daniela Rossi3Vincenzo Sorrentino4Luca Bini5Functional Proteomics Lab., Department Life Sciences, University of Siena, Via Aldo Moro 2, 53100 Siena, ItalyFunctional Proteomics Lab., Department Life Sciences, University of Siena, Via Aldo Moro 2, 53100 Siena, ItalyFunctional Proteomics Lab., Department Life Sciences, University of Siena, Via Aldo Moro 2, 53100 Siena, ItalyDepartment of Molecular and Developmental Medicine, University of Siena, Via Aldo Moro 2, 53100 Siena, ItalyDepartment of Molecular and Developmental Medicine, University of Siena, Via Aldo Moro 2, 53100 Siena, ItalyFunctional Proteomics Lab., Department Life Sciences, University of Siena, Via Aldo Moro 2, 53100 Siena, ItalyMuscle 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.https://www.mdpi.com/2227-7382/12/4/27protein denaturationmuscle tissuetwo-dimensional electrophoresismass spectrometrymyopathies |
| spellingShingle | Lorenza Vantaggiato Claudia Landi Enxhi Shaba Daniela Rossi Vincenzo Sorrentino Luca Bini Protein Extraction Methods Suitable for Muscle Tissue Proteomic Analysis Proteomes protein denaturation muscle tissue two-dimensional electrophoresis mass spectrometry myopathies |
| title | Protein Extraction Methods Suitable for Muscle Tissue Proteomic Analysis |
| title_full | Protein Extraction Methods Suitable for Muscle Tissue Proteomic Analysis |
| title_fullStr | Protein Extraction Methods Suitable for Muscle Tissue Proteomic Analysis |
| title_full_unstemmed | Protein Extraction Methods Suitable for Muscle Tissue Proteomic Analysis |
| title_short | Protein Extraction Methods Suitable for Muscle Tissue Proteomic Analysis |
| title_sort | protein extraction methods suitable for muscle tissue proteomic analysis |
| topic | protein denaturation muscle tissue two-dimensional electrophoresis mass spectrometry myopathies |
| url | https://www.mdpi.com/2227-7382/12/4/27 |
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