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Characterization of Contractile Proteins from Skeletal Muscle Using Gel-Based Top-Down Proteomics
The mass spectrometric analysis of skeletal muscle proteins has used both peptide-centric and protein-focused approaches. The term ‘top-down proteomics’ is often used in relation to studying purified proteoforms and their post-translational modifications. Two-dimensional gel electrophoresis, in comb...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6631179/ https://www.ncbi.nlm.nih.gov/pubmed/31226838 http://dx.doi.org/10.3390/proteomes7020025 |
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author | Dowling, Paul Zweyer, Margit Swandulla, Dieter Ohlendieck, Kay |
author_facet | Dowling, Paul Zweyer, Margit Swandulla, Dieter Ohlendieck, Kay |
author_sort | Dowling, Paul |
collection | PubMed |
description | The mass spectrometric analysis of skeletal muscle proteins has used both peptide-centric and protein-focused approaches. The term ‘top-down proteomics’ is often used in relation to studying purified proteoforms and their post-translational modifications. Two-dimensional gel electrophoresis, in combination with peptide generation for the identification and characterization of intact proteoforms being present in two-dimensional spots, plays a critical role in specific applications of top-down proteomics. A decisive bioanalytical advantage of gel-based and top-down approaches is the initial bioanalytical focus on intact proteins, which usually enables the swift identification and detailed characterisation of specific proteoforms. In this review, we describe the usage of two-dimensional gel electrophoretic top-down proteomics and related approaches for the systematic analysis of key components of the contractile apparatus, with a special focus on myosin heavy and light chains and their associated regulatory proteins. The detailed biochemical analysis of proteins belonging to the thick and thin skeletal muscle filaments has decisively improved our biochemical understanding of structure-function relationships within the contractile apparatus. Gel-based and top-down proteomics has clearly established a variety of slow and fast isoforms of myosin, troponin and tropomyosin as excellent markers of fibre type specification and dynamic muscle transition processes. |
format | Online Article Text |
id | pubmed-6631179 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-66311792019-08-19 Characterization of Contractile Proteins from Skeletal Muscle Using Gel-Based Top-Down Proteomics Dowling, Paul Zweyer, Margit Swandulla, Dieter Ohlendieck, Kay Proteomes Review The mass spectrometric analysis of skeletal muscle proteins has used both peptide-centric and protein-focused approaches. The term ‘top-down proteomics’ is often used in relation to studying purified proteoforms and their post-translational modifications. Two-dimensional gel electrophoresis, in combination with peptide generation for the identification and characterization of intact proteoforms being present in two-dimensional spots, plays a critical role in specific applications of top-down proteomics. A decisive bioanalytical advantage of gel-based and top-down approaches is the initial bioanalytical focus on intact proteins, which usually enables the swift identification and detailed characterisation of specific proteoforms. In this review, we describe the usage of two-dimensional gel electrophoretic top-down proteomics and related approaches for the systematic analysis of key components of the contractile apparatus, with a special focus on myosin heavy and light chains and their associated regulatory proteins. The detailed biochemical analysis of proteins belonging to the thick and thin skeletal muscle filaments has decisively improved our biochemical understanding of structure-function relationships within the contractile apparatus. Gel-based and top-down proteomics has clearly established a variety of slow and fast isoforms of myosin, troponin and tropomyosin as excellent markers of fibre type specification and dynamic muscle transition processes. MDPI 2019-06-20 /pmc/articles/PMC6631179/ /pubmed/31226838 http://dx.doi.org/10.3390/proteomes7020025 Text en © 2019 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Review Dowling, Paul Zweyer, Margit Swandulla, Dieter Ohlendieck, Kay Characterization of Contractile Proteins from Skeletal Muscle Using Gel-Based Top-Down Proteomics |
title | Characterization of Contractile Proteins from Skeletal Muscle Using Gel-Based Top-Down Proteomics |
title_full | Characterization of Contractile Proteins from Skeletal Muscle Using Gel-Based Top-Down Proteomics |
title_fullStr | Characterization of Contractile Proteins from Skeletal Muscle Using Gel-Based Top-Down Proteomics |
title_full_unstemmed | Characterization of Contractile Proteins from Skeletal Muscle Using Gel-Based Top-Down Proteomics |
title_short | Characterization of Contractile Proteins from Skeletal Muscle Using Gel-Based Top-Down Proteomics |
title_sort | characterization of contractile proteins from skeletal muscle using gel-based top-down proteomics |
topic | Review |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6631179/ https://www.ncbi.nlm.nih.gov/pubmed/31226838 http://dx.doi.org/10.3390/proteomes7020025 |
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