Age- and Activity-Related Differences in the Abundance of Myosin Essential and Regulatory Light Chains in Human Muscle
Traditional methods for phenotyping skeletal muscle (e.g., immunohistochemistry) are labor-intensive and ill-suited to multixplex analysis, i.e., assays must be performed in a series. Addressing these concerns represents a largely unmet research need but more comprehensive parallel analysis of myofi...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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MDPI AG
2016-04-01
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| Series: | Proteomes |
| Subjects: | |
| Online Access: | http://www.mdpi.com/2227-7382/4/2/15 |
| _version_ | 1811184762440646656 |
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| author | James N. Cobley Zulezwan Ab. Malik James P. Morton Graeme L. Close Ben J. Edwards Jatin G. Burniston |
| author_facet | James N. Cobley Zulezwan Ab. Malik James P. Morton Graeme L. Close Ben J. Edwards Jatin G. Burniston |
| author_sort | James N. Cobley |
| collection | DOAJ |
| description | Traditional methods for phenotyping skeletal muscle (e.g., immunohistochemistry) are labor-intensive and ill-suited to multixplex analysis, i.e., assays must be performed in a series. Addressing these concerns represents a largely unmet research need but more comprehensive parallel analysis of myofibrillar proteins could advance knowledge regarding age- and activity-dependent changes in human muscle. We report a label-free, semi-automated and time efficient LC-MS proteomic workflow for phenotyping the myofibrillar proteome. Application of this workflow in old and young as well as trained and untrained human skeletal muscle yielded several novel observations that were subsequently verified by multiple reaction monitoring (MRM). We report novel data demonstrating that human ageing is associated with lesser myosin light chain 1 content and greater myosin light chain 3 content, consistent with an age-related reduction in type II muscle fibers. We also disambiguate conflicting data regarding myosin regulatory light chain, revealing that age-related changes in this protein more closely reflect physical activity status than ageing per se. This finding reinforces the need to control for physical activity levels when investigating the natural process of ageing. Taken together, our data confirm and extend knowledge regarding age- and activity-related phenotypes. In addition, the MRM transitions described here provide a methodological platform that can be fine-tuned to suite multiple research needs and thus advance myofibrillar phenotyping. |
| first_indexed | 2024-04-11T13:19:00Z |
| format | Article |
| id | doaj.art-43ad81c85b7443ca8b47a115ff2ac32e |
| institution | Directory Open Access Journal |
| issn | 2227-7382 |
| language | English |
| last_indexed | 2024-04-11T13:19:00Z |
| publishDate | 2016-04-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Proteomes |
| spelling | doaj.art-43ad81c85b7443ca8b47a115ff2ac32e2022-12-22T04:22:18ZengMDPI AGProteomes2227-73822016-04-01421510.3390/proteomes4020015proteomes4020015Age- and Activity-Related Differences in the Abundance of Myosin Essential and Regulatory Light Chains in Human MuscleJames N. Cobley0Zulezwan Ab. Malik1James P. Morton2Graeme L. Close3Ben J. Edwards4Jatin G. Burniston5Division of Sport and Exercise Science, Abertay University, 40 Bell Street, Kydd Building, Dundee DD1 1HG, UKFaculty of Sport Science and Coaching, Universiti Pendidikan Sultan Idris, Tanjung Malim, Perak 35900, MalaysiaResearch Institute for Sport and Exercise Sciences, Liverpool John Moores University, Liverpool L3 3AF, UKResearch Institute for Sport and Exercise Sciences, Liverpool John Moores University, Liverpool L3 3AF, UKResearch Institute for Sport and Exercise Sciences, Liverpool John Moores University, Liverpool L3 3AF, UKResearch Institute for Sport and Exercise Sciences, Liverpool John Moores University, Liverpool L3 3AF, UKTraditional methods for phenotyping skeletal muscle (e.g., immunohistochemistry) are labor-intensive and ill-suited to multixplex analysis, i.e., assays must be performed in a series. Addressing these concerns represents a largely unmet research need but more comprehensive parallel analysis of myofibrillar proteins could advance knowledge regarding age- and activity-dependent changes in human muscle. We report a label-free, semi-automated and time efficient LC-MS proteomic workflow for phenotyping the myofibrillar proteome. Application of this workflow in old and young as well as trained and untrained human skeletal muscle yielded several novel observations that were subsequently verified by multiple reaction monitoring (MRM). We report novel data demonstrating that human ageing is associated with lesser myosin light chain 1 content and greater myosin light chain 3 content, consistent with an age-related reduction in type II muscle fibers. We also disambiguate conflicting data regarding myosin regulatory light chain, revealing that age-related changes in this protein more closely reflect physical activity status than ageing per se. This finding reinforces the need to control for physical activity levels when investigating the natural process of ageing. Taken together, our data confirm and extend knowledge regarding age- and activity-related phenotypes. In addition, the MRM transitions described here provide a methodological platform that can be fine-tuned to suite multiple research needs and thus advance myofibrillar phenotyping.http://www.mdpi.com/2227-7382/4/2/15ageinghuman skeletal musclemyosin heavy chainmyosin light chainselective reaction monitoring |
| spellingShingle | James N. Cobley Zulezwan Ab. Malik James P. Morton Graeme L. Close Ben J. Edwards Jatin G. Burniston Age- and Activity-Related Differences in the Abundance of Myosin Essential and Regulatory Light Chains in Human Muscle Proteomes ageing human skeletal muscle myosin heavy chain myosin light chain selective reaction monitoring |
| title | Age- and Activity-Related Differences in the Abundance of Myosin Essential and Regulatory Light Chains in Human Muscle |
| title_full | Age- and Activity-Related Differences in the Abundance of Myosin Essential and Regulatory Light Chains in Human Muscle |
| title_fullStr | Age- and Activity-Related Differences in the Abundance of Myosin Essential and Regulatory Light Chains in Human Muscle |
| title_full_unstemmed | Age- and Activity-Related Differences in the Abundance of Myosin Essential and Regulatory Light Chains in Human Muscle |
| title_short | Age- and Activity-Related Differences in the Abundance of Myosin Essential and Regulatory Light Chains in Human Muscle |
| title_sort | age and activity related differences in the abundance of myosin essential and regulatory light chains in human muscle |
| topic | ageing human skeletal muscle myosin heavy chain myosin light chain selective reaction monitoring |
| url | http://www.mdpi.com/2227-7382/4/2/15 |
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