Dowling, Paul and Zweyer, Margit and Swandulla, Dieter and Ohlendieck, Kay (2019) Characterization of Contractile Proteins from Skeletal Muscle Using Gel-Based Top-Down Proteomics. Proteomes, 7 (2). ISSN 2227-7382

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Abstract

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.

Item Type:	Article
Additional Information:	© 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/). Cite as: Dowling, P.; Zweyer, M.; Swandulla, D.; Ohlendieck, K. Characterization of Contractile Proteins from Skeletal Muscle Using Gel-Based Top-Down Proteomics. Proteomes 2019, 7, 25. Funding: Research was supported by grants from Muscular Dystrophy Ireland and the Irish Health Research Board (HRB/MRCG-2016-20) and the Human Health Research Institute of Maynooth University
Keywords:	actin; mass spectrometry; muscle proteomics; myosin; top-down proteomics; tropomyosin; troponin; two-dimensional gel electrophoresis; skeletal muscle;
Academic Unit:	Faculty of Science and Engineering > Biology
Item ID:	13572
Identification Number:	https://doi.org/10.3390/proteomes7020025
Depositing User:	Paul Dowling
Date Deposited:	18 Nov 2020 16:17
Journal or Publication Title:	Proteomes
Publisher:	MDPI
Refereed:	Yes
Funders:	Muscular Dystrophy Ireland, Health Research Board (HRB), Human Health Institute, Maynooth University
URI:	Publisher
Use Licence:	This item is available under a Creative Commons Attribution Non Commercial Share Alike Licence (CC BY-NC-SA). Details of this licence are available here

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