TY - JOUR A1 - Lang, Franziska A1 - Aravamudhan, Sriram A1 - Nolte, Hendrik A1 - Türk, Clara A1 - Hölper, Soraya A1 - Müller, Stefan A1 - Günther, Stefan A1 - Blaauw, Bert A1 - Braun, Thomas A1 - Krüger, Marcus T1 - Dynamic changes in the mouse skeletal muscle proteome during denervation-induced atrophy T2 - Disease models & mechanisms N2 - Loss of neuronal stimulation enhances protein breakdown and reduces protein synthesis, causing rapid loss of muscle mass. To elucidate the pathophysiological adaptations that occur in atrophying muscles, we used stable isotope labelling and mass spectrometry to quantify protein expression changes accurately during denervation-induced atrophy after sciatic nerve section in the mouse gastrocnemius muscle. Additionally, mice were fed a stable isotope labelling of amino acids in cell culture (SILAC) diet containing 13C6-lysine for 4, 7 or 11 days to calculate relative levels of protein synthesis in denervated and control muscles. Ubiquitin remnant peptides (K-ε-GG) were profiled by immunoaffinity enrichment to identify potential substrates of the ubiquitin-proteasomal pathway. Of the 4279 skeletal muscle proteins quantified, 850 were differentially expressed significantly within 2 weeks after denervation compared with control muscles. Moreover, pulse labelling identified Lys6 incorporation in 4786 proteins, of which 43 had differential Lys6 incorporation between control and denervated muscle. Enrichment of diglycine remnants identified 2100 endogenous ubiquitination sites and revealed a metabolic and myofibrillar protein diglycine signature, including myosin heavy chains, myomesins and titin, during denervation. Comparative analysis of these proteomic data sets with known atrogenes using a random forest approach identified 92 proteins subject to atrogene-like regulation that have not previously been associated directly with denervation-induced atrophy. Comparison of protein synthesis and proteomic data indicated that upregulation of specific proteins in response to denervation is mainly achieved by protein stabilization. This study provides the first integrated analysis of protein expression, synthesis and ubiquitin signatures during muscular atrophy in a living animal. KW - Muscle atrophy KW - Denervation KW - Pulsed SILAC KW - Ubiquitination KW - Random forest Y1 - 2017 UR - http://publikationen.ub.uni-frankfurt.de/frontdoor/index/index/docId/45073 UR - https://nbn-resolving.org/urn:nbn:de:hebis:30:3-450736 SN - 1754-8411 SN - 1754-8403 N1 - This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution and reproduction in any medium provided that the original work is properly attributed VL - 10 IS - 7 SP - 881 EP - 896 PB - Company of Biologists Limited CY - Cambridge ER -