Kallabis, Sebastian, Abraham, Lena, Mueller, Stefan, Dzialas, Verena, Tuerk, Clara, Wiederstein, Janica Lea, Bock, Theresa, Nolte, Hendrik, Nogara, Leonardo, Blaauw, Bert, Braun, Thomas and Krueger, Marcus (2020). High-throughput proteomics fiber typing (ProFiT) for comprehensive characterization of single skeletal muscle fibers. Skeletal Muscle, 10 (1). LONDON: BMC. ISSN 2044-5040

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Abstract

Background Skeletal muscles are composed of a heterogeneous collection of fiber types with different physiological adaption in response to a stimulus and disease-related conditions. Each fiber has a specific molecular expression of myosin heavy chain molecules (MyHC). So far, MyHCs are currently the best marker proteins for characterization of individual fiber types, and several proteome profiling studies have helped to dissect the molecular signature of whole muscles and individual fibers. Methods Herein, we describe a mass spectrometric workflow to measure skeletal muscle fiber type-specific proteomes. To bypass the limited quantities of protein in single fibers, we developed a Proteomics high-throughput fiber typing (ProFiT) approach enabling profiling of MyHC in single fibers. Aliquots of protein extracts from separated muscle fibers were subjected to capillary LC-MS gradients to profile MyHC isoforms in a 96-well format. Muscle fibers with the same MyHC protein expression were pooled and subjected to proteomic, pulsed-SILAC, and phosphoproteomic analysis. Results Our fiber type-specific quantitative proteome analysis confirmed the distribution of fiber types in the soleus muscle, substantiates metabolic adaptions in oxidative and glycolytic fibers, and highlighted significant differences between the proteomes of type IIb fibers from different muscle groups, including a differential expression of desmin and actinin-3. A detailed map of the Lys-6 incorporation rates in muscle fibers showed an increased turnover of slow fibers compared to fast fibers. In addition, labeling of mitochondrial respiratory chain complexes revealed a broad range of Lys-6 incorporation rates, depending on the localization of the subunits within distinct complexes. Conclusion Overall, the ProFiT approach provides a versatile tool to rapidly characterize muscle fibers and obtain fiber-specific proteomes for different muscle groups.

Item Type: Journal Article
Creators:
CreatorsEmailORCIDORCID Put Code
Kallabis, SebastianUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Abraham, LenaUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Mueller, StefanUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Dzialas, VerenaUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Tuerk, ClaraUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Wiederstein, Janica LeaUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Bock, TheresaUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Nolte, HendrikUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Nogara, LeonardoUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Blaauw, BertUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Braun, ThomasUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
Krueger, MarcusUNSPECIFIEDUNSPECIFIEDUNSPECIFIED
URN: urn:nbn:de:hbz:38-340417
DOI: 10.1186/s13395-020-00226-5
Journal or Publication Title: Skeletal Muscle
Volume: 10
Number: 1
Date: 2020
Publisher: BMC
Place of Publication: LONDON
ISSN: 2044-5040
Language: English
Faculty: Unspecified
Divisions: Unspecified
Subjects: no entry
Uncontrolled Keywords:
KeywordsLanguage
LIGHT-CHAIN PHOSPHORYLATION; COMPLEX I; ENCODED SUBUNITS; GENE-EXPRESSION; MYOSIN ISOFORMS; MITOCHONDRIAL; TURNOVER; ACTIVATION; MECHANISM; DYNAMICSMultiple languages
Cell BiologyMultiple languages
URI: http://kups.ub.uni-koeln.de/id/eprint/34041

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