Purification and quantitative proteomic analysis of cell bodies and protrusions
Summary: Actin-rich protrusions are membrane extensions generated by actin polymerization that drive mesenchymal-like cell migration. Characterization of protrusions proteome is crucial for understanding their function. We present a complete step-by-step protocol based on microporous filter-based fr...
| Main Authors: | , |
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| Format: | Article |
| Language: | English |
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Elsevier
2021-06-01
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| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166721001696 |
| _version_ | 1819102588364128256 |
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| author | Maria Dermit Faraz K. Mardakheh |
| author_facet | Maria Dermit Faraz K. Mardakheh |
| author_sort | Maria Dermit |
| collection | DOAJ |
| description | Summary: Actin-rich protrusions are membrane extensions generated by actin polymerization that drive mesenchymal-like cell migration. Characterization of protrusions proteome is crucial for understanding their function. We present a complete step-by-step protocol based on microporous filter-based fractionation of protrusive cellular domains coupled with sample preparation for quantitative proteomics, mass spectrometric data acquisition, and data analysis. This protocol enables purification, quantification, and analysis of the distribution of proteins present in protrusions and cell bodies.For complete details on the use and execution of this protocol, please refer to Dermit et al. (2020). |
| first_indexed | 2024-12-22T01:36:57Z |
| format | Article |
| id | doaj.art-52f1153d0c784b16bfda1422a13f12d5 |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-12-22T01:36:57Z |
| publishDate | 2021-06-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-52f1153d0c784b16bfda1422a13f12d52022-12-21T18:43:20ZengElsevierSTAR Protocols2666-16672021-06-0122100462Purification and quantitative proteomic analysis of cell bodies and protrusionsMaria Dermit0Faraz K. Mardakheh1Centre for Cancer Cell and Molecular Biology, Barts Cancer Institute, Queen Mary University of London, Charterhouse Square, London EC1M 6BQ, UK; Corresponding authorCentre for Cancer Cell and Molecular Biology, Barts Cancer Institute, Queen Mary University of London, Charterhouse Square, London EC1M 6BQ, UK; Corresponding authorSummary: Actin-rich protrusions are membrane extensions generated by actin polymerization that drive mesenchymal-like cell migration. Characterization of protrusions proteome is crucial for understanding their function. We present a complete step-by-step protocol based on microporous filter-based fractionation of protrusive cellular domains coupled with sample preparation for quantitative proteomics, mass spectrometric data acquisition, and data analysis. This protocol enables purification, quantification, and analysis of the distribution of proteins present in protrusions and cell bodies.For complete details on the use and execution of this protocol, please refer to Dermit et al. (2020).http://www.sciencedirect.com/science/article/pii/S2666166721001696BioinformaticsCell biologyCell separation/fractionationProteomicsMass spectrometry |
| spellingShingle | Maria Dermit Faraz K. Mardakheh Purification and quantitative proteomic analysis of cell bodies and protrusions STAR Protocols Bioinformatics Cell biology Cell separation/fractionation Proteomics Mass spectrometry |
| title | Purification and quantitative proteomic analysis of cell bodies and protrusions |
| title_full | Purification and quantitative proteomic analysis of cell bodies and protrusions |
| title_fullStr | Purification and quantitative proteomic analysis of cell bodies and protrusions |
| title_full_unstemmed | Purification and quantitative proteomic analysis of cell bodies and protrusions |
| title_short | Purification and quantitative proteomic analysis of cell bodies and protrusions |
| title_sort | purification and quantitative proteomic analysis of cell bodies and protrusions |
| topic | Bioinformatics Cell biology Cell separation/fractionation Proteomics Mass spectrometry |
| url | http://www.sciencedirect.com/science/article/pii/S2666166721001696 |
| work_keys_str_mv | AT mariadermit purificationandquantitativeproteomicanalysisofcellbodiesandprotrusions AT farazkmardakheh purificationandquantitativeproteomicanalysisofcellbodiesandprotrusions |