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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...

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Autores principales: Dowling, Paul, Zweyer, Margit, Swandulla, Dieter, Ohlendieck, Kay
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2019
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.
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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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