Protein profile of fiber types in human skeletal muscle: a single-fiber proteomics study

Abstract Background Human skeletal muscle is composed of three major fiber types, referred to as type 1, 2A, and 2X fibers. This heterogeneous cellular composition complicates the interpretation of studies based on whole skeletal muscle lysate. A single-fiber proteomics approach is required to obtai...

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Autores principales:	Marta Murgia, Leonardo Nogara, Martina Baraldo, Carlo Reggiani, Matthias Mann, Stefano Schiaffino
Formato:	article
Lenguaje:	EN
Publicado:	BMC 2021
Materias:	Human skeletal muscle Muscle fiber types Single-fiber proteomics Mass spectrometry Diseases of the musculoskeletal system RC925-935
Acceso en línea:	https://doaj.org/article/9a2d90d6736b4e079c06502c1b989d35
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spelling	oai:doaj.org-article:9a2d90d6736b4e079c06502c1b989d352021-11-07T12:06:33ZProtein profile of fiber types in human skeletal muscle: a single-fiber proteomics study10.1186/s13395-021-00279-02044-5040https://doaj.org/article/9a2d90d6736b4e079c06502c1b989d352021-11-01T00:00:00Zhttps://doi.org/10.1186/s13395-021-00279-0https://doaj.org/toc/2044-5040Abstract Background Human skeletal muscle is composed of three major fiber types, referred to as type 1, 2A, and 2X fibers. This heterogeneous cellular composition complicates the interpretation of studies based on whole skeletal muscle lysate. A single-fiber proteomics approach is required to obtain a fiber-type resolved quantitative information on skeletal muscle pathophysiology. Methods Single fibers were dissected from vastus lateralis muscle biopsies of young adult males and processed for mass spectrometry-based single-fiber proteomics. We provide and analyze a resource dataset based on relatively pure fibers, containing at least 80% of either MYH7 (marker of slow type 1 fibers), MYH2 (marker of fast 2A fibers), or MYH1 (marker of fast 2X fibers). Results In a dataset of more than 3800 proteins detected by single-fiber proteomics, we selected 404 proteins showing a statistically significant difference among fiber types. We identified numerous type 1 or 2X fiber type–specific protein markers, defined as proteins present at 3-fold or higher levels in these compared to other fiber types. In contrast, we could detect only two 2A-specific protein markers in addition to MYH2. We observed three other major patterns: proteins showing a differential distribution according to the sequence 1 > 2A > 2X or 2X > 2A > 1 and type 2–specific proteins expressed in 2A and 2X fibers at levels 3 times greater than in type 1 fibers. In addition to precisely quantifying known fiber type–specific protein patterns, our study revealed several novel features of fiber type specificity, including the selective enrichment of components of the dystrophin and integrin complexes, as well as microtubular proteins, in type 2X fibers. The fiber type–specific distribution of some selected proteins revealed by proteomics was validated by immunofluorescence analyses with specific antibodies. Conclusion We here show that numerous muscle proteins, including proteins whose function is unknown, are selectively enriched in specific fiber types, pointing to potential implications in muscle pathophysiology. This reinforces the notion that single-fiber proteomics, together with recently developed approaches to single-cell proteomics, will be instrumental to explore and quantify muscle cell heterogeneity.Marta MurgiaLeonardo NogaraMartina BaraldoCarlo ReggianiMatthias MannStefano SchiaffinoBMCarticleHuman skeletal muscleMuscle fiber typesSingle-fiber proteomicsMass spectrometryDiseases of the musculoskeletal systemRC925-935ENSkeletal Muscle, Vol 11, Iss 1, Pp 1-19 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Human skeletal muscle Muscle fiber types Single-fiber proteomics Mass spectrometry Diseases of the musculoskeletal system RC925-935
spellingShingle	Human skeletal muscle Muscle fiber types Single-fiber proteomics Mass spectrometry Diseases of the musculoskeletal system RC925-935 Marta Murgia Leonardo Nogara Martina Baraldo Carlo Reggiani Matthias Mann Stefano Schiaffino Protein profile of fiber types in human skeletal muscle: a single-fiber proteomics study
description	Abstract Background Human skeletal muscle is composed of three major fiber types, referred to as type 1, 2A, and 2X fibers. This heterogeneous cellular composition complicates the interpretation of studies based on whole skeletal muscle lysate. A single-fiber proteomics approach is required to obtain a fiber-type resolved quantitative information on skeletal muscle pathophysiology. Methods Single fibers were dissected from vastus lateralis muscle biopsies of young adult males and processed for mass spectrometry-based single-fiber proteomics. We provide and analyze a resource dataset based on relatively pure fibers, containing at least 80% of either MYH7 (marker of slow type 1 fibers), MYH2 (marker of fast 2A fibers), or MYH1 (marker of fast 2X fibers). Results In a dataset of more than 3800 proteins detected by single-fiber proteomics, we selected 404 proteins showing a statistically significant difference among fiber types. We identified numerous type 1 or 2X fiber type–specific protein markers, defined as proteins present at 3-fold or higher levels in these compared to other fiber types. In contrast, we could detect only two 2A-specific protein markers in addition to MYH2. We observed three other major patterns: proteins showing a differential distribution according to the sequence 1 > 2A > 2X or 2X > 2A > 1 and type 2–specific proteins expressed in 2A and 2X fibers at levels 3 times greater than in type 1 fibers. In addition to precisely quantifying known fiber type–specific protein patterns, our study revealed several novel features of fiber type specificity, including the selective enrichment of components of the dystrophin and integrin complexes, as well as microtubular proteins, in type 2X fibers. The fiber type–specific distribution of some selected proteins revealed by proteomics was validated by immunofluorescence analyses with specific antibodies. Conclusion We here show that numerous muscle proteins, including proteins whose function is unknown, are selectively enriched in specific fiber types, pointing to potential implications in muscle pathophysiology. This reinforces the notion that single-fiber proteomics, together with recently developed approaches to single-cell proteomics, will be instrumental to explore and quantify muscle cell heterogeneity.
format	article
author	Marta Murgia Leonardo Nogara Martina Baraldo Carlo Reggiani Matthias Mann Stefano Schiaffino
author_facet	Marta Murgia Leonardo Nogara Martina Baraldo Carlo Reggiani Matthias Mann Stefano Schiaffino
author_sort	Marta Murgia
title	Protein profile of fiber types in human skeletal muscle: a single-fiber proteomics study
title_short	Protein profile of fiber types in human skeletal muscle: a single-fiber proteomics study
title_full	Protein profile of fiber types in human skeletal muscle: a single-fiber proteomics study
title_fullStr	Protein profile of fiber types in human skeletal muscle: a single-fiber proteomics study
title_full_unstemmed	Protein profile of fiber types in human skeletal muscle: a single-fiber proteomics study
title_sort	protein profile of fiber types in human skeletal muscle: a single-fiber proteomics study
publisher	BMC
publishDate	2021
url	https://doaj.org/article/9a2d90d6736b4e079c06502c1b989d35
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Protein profile of fiber types in human skeletal muscle: a single-fiber proteomics study

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