{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,27]],"date-time":"2025-10-27T20:43:55Z","timestamp":1761597835211},"reference-count":41,"publisher":"Wiley","issue":"5","license":[{"start":{"date-parts":[[2017,7,3]],"date-time":"2017-07-03T00:00:00Z","timestamp":1499040000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100004243","name":"Prinses Beatrix Spierfonds","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100004243","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["J cachexia sarcopenia muscle"],"published-print":{"date-parts":[[2017,10]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:sec><jats:title>Background<\/jats:title><jats:p>Protein homeostasis, primarily regulated by the ubiquitin\u2013proteasome system is crucial for proper function of cells. In tissues of post\u2010mitotic cells, the impaired ubiquitin\u2013proteasome system is found in a wide range of neuromuscular disorders. Activity\u2010based probes (ABPs) measure proteasomal proteolytic subunits and can be used to report protein homeostasis. Despite the crucial role of the proteasome in neuromuscular pathologies, ABPs were not employed in muscle cells and tissues, and measurement of proteasomal activity was carried out <jats:italic>in vitro<\/jats:italic> using low\u2010throughput procedures.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>We screened six ABPs for specific application in muscle cell culture using high throughput call\u2010based imaging procedures. We then determined an <jats:italic>in situ<\/jats:italic> proteasomal activity in myofibers of muscle cryosections.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>We demonstrate that LWA300, a pan\u2010reactive proteasomal probe, is most suitable to report proteasomal activity in muscle cells using cell\u2010based bio\u2010imaging. We found that proteasomal activity is two\u2010fold and three\u2010fold enhanced in fused muscle cell culture compared with non\u2010fused cells. Moreover, we found that proteasomal activity can discriminate between muscles. Across muscles, a relative higher proteasomal activity was found in hybrid myofibers whereas fast\u2010twitch myofibers displayed lower activity.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusions<\/jats:title><jats:p>Our study demonstrates that proteasomal activity differ between muscles and between myofiber types. We suggest that ABPs can be used to report disease progression and treatment efficacy.<\/jats:p><\/jats:sec>","DOI":"10.1002\/jcsm.12211","type":"journal-article","created":{"date-parts":[[2017,7,4]],"date-time":"2017-07-04T13:17:52Z","timestamp":1499174272000},"page":"798-807","update-policy":"http:\/\/dx.doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["Proteasomal activity\u2010based probes mark protein homeostasis in muscles"],"prefix":"10.1002","volume":"8","author":[{"given":"Vered","family":"Raz","sequence":"first","affiliation":[{"name":"Department of Human Genetics LUMC  Leiden The Netherlands"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yotam","family":"Raz","sequence":"additional","affiliation":[{"name":"Department of Human Genetics LUMC  Leiden The 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