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Tytuł:
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Comparison of extracellular matrix enrichment protocols for the improved characterization of the skin matrisome by mass spectrometry.
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Autorzy:
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Dussoyer M; University of Lyon, CNRS, Tissue Biology and Therapeutic Engineering Laboratory, LBTI, UMR5305, F-69367 Lyon, France.
Page A; University of Lyon, INSERM, ENS Lyon, CNRS, Protein Science Facility, SFR BioSciences, UAR3444/US8, F-69366 Lyon, France.
Delolme F; University of Lyon, INSERM, ENS Lyon, CNRS, Protein Science Facility, SFR BioSciences, UAR3444/US8, F-69366 Lyon, France.
Rousselle P; University of Lyon, CNRS, Tissue Biology and Therapeutic Engineering Laboratory, LBTI, UMR5305, F-69367 Lyon, France.
Nyström A; Department of Clinical Dermatology/Medical Center, University of Freiburg, Freiburg, Germany; Freiburg Institute for Advanced Studies (FRIAS), University of Freiburg, Freiburg, Germany.
Moali C; University of Lyon, CNRS, Tissue Biology and Therapeutic Engineering Laboratory, LBTI, UMR5305, F-69367 Lyon, France. Electronic address: .
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Źródło:
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Journal of proteomics [J Proteomics] 2022 Jan 16; Vol. 251, pp. 104397. Date of Electronic Publication: 2021 Oct 20.
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Typ publikacji:
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Comparative Study; Journal Article; Research Support, Non-U.S. Gov't
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Język:
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English
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Imprint Name(s):
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Original Publication: Amsterdam : Elsevier
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MeSH Terms:
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Extracellular Matrix*/metabolism
Proteomics*/methods
Animals ; Extracellular Matrix Proteins/analysis ; Mass Spectrometry ; Mice ; Skin/metabolism
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Contributed Indexing:
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Keywords: Decellularization; Extracellular matrix; Matrisome; Mouse; Proteomics; Skin
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Substance Nomenclature:
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0 (Extracellular Matrix Proteins)
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Entry Date(s):
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Date Created: 20211022 Date Completed: 20220225 Latest Revision: 20220531
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Update Code:
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20240105
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DOI:
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10.1016/j.jprot.2021.104397
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PMID:
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34678517
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A striking feature of skin organization is that the extracellular matrix (ECM) occupies a larger volume than the cells. Skin ECM also directly contributes to aging and most cutaneous diseases. In recent years, specific ECM enrichment protocols combined with in silico approaches allowed the proteomic description of the matrisome of various organs and tumor samples. Nevertheless, the skin matrisome remains under-studied and protocols allowing the efficient recovery of the diverse ECM found in skin are still to be described. Here, we compared four protocols allowing the enrichment of ECM proteins from adult mouse back skin and found that all protocols led to a significant enrichment (up to 65%) of matrisome proteins when compared to total skin lysates. The protocols based on decellularization and solubility profiling gave the best results in terms of numbers of proteins identified and confirmed that skin matrisome proteins exhibit very diverse solubility and abundance profiles. We also report the first description of the skin matrisome of healthy adult mice that includes 236 proteins comprising 95 core matrisome proteins and 141 associated matrisome proteins. These results provide a reliable basis for future characterizations of skin ECM proteins and their dysregulations in disease-specific contexts. SIGNIFICANCE: Extracellular matrix proteins are key players in skin physiopathology and have been involved in several diseases such as genetic disorders, wound healing defects, scleroderma and skin carcinoma. However, skin ECM proteins are numerous, diverse and challenging to analyze by mass spectrometry due to the multiplicity of their post-translational modifications and to the heterogeneity of their solubility profiles. Here, we performed the thorough evaluation of four ECM enrichment protocols compatible with the proteomic analysis of mouse back skin and provide the first description of the adult mouse skin matrisome in homeostasis conditions. Our work will greatly facilitate the future characterization of skin ECM alterations in preclinical mouse models and will inspire new optimizations to analyze the skin matrisome of other species and of human clinical samples.
(Copyright © 2021 Elsevier B.V. All rights reserved.)
