{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,11,11]],"date-time":"2025-11-11T22:20:37Z","timestamp":1762899637016,"version":"build-2065373602"},"reference-count":45,"publisher":"MDPI AG","issue":"11","license":[{"start":{"date-parts":[[2018,11,13]],"date-time":"2018-11-13T00:00:00Z","timestamp":1542067200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Methylmalonic acidemias (MMAs) are inborn errors of metabolism due to the deficient activity of methylmalonyl-CoA mutase (MUT). MUT catalyzes the formation of succinyl-CoA from methylmalonyl-CoA, produced from propionyl-CoA catabolism and derived from odd chain fatty acids \u03b2-oxidation, cholesterol, and branched-chain amino acids degradation. Increased methylmalonyl-CoA levels allow for the presymptomatic diagnosis of the disease, even though no approved therapies exist. MMA patients show hyperammonemia, ketoacidosis, lethargy, respiratory distress, cognitive impairment, and hepatomegaly. The long-term consequences concern neurologic damage and terminal kidney failure, with little chance of survival. The cellular pathways affected by MUT deficiency were investigated using a quantitative proteomics approach on a cellular model of MUT knockdown. Currently, a consistent reduction of the MUT protein expression was obtained in the neuroblastoma cell line (SH-SY5Y) by using small-interfering RNA (siRNA) directed against an MUT transcript (MUT siRNA). The MUT absence did not affect the cell viability and apoptotic process in SH-SY5Y. In the present study, we evaluate and quantify the alterations in the protein expression profile as a consequence of MUT-silencing by a mass spectrometry-based label-free quantitative analysis, using two different quantitative strategies. Both quantitative methods allowed us to observe that the expression of the proteins involved in mitochondrial oxido-reductive homeostasis balance was affected by MUT deficiency. The alterated functional mitochondrial activity was observed in siRNA_MUT cells cultured with a propionate-supplemented medium. Finally, alterations in the levels of proteins involved in the metabolic pathways, like carbohydrate metabolism and lipid metabolism, were found.<\/jats:p>","DOI":"10.3390\/ijms19113580","type":"journal-article","created":{"date-parts":[[2018,11,14]],"date-time":"2018-11-14T02:42:41Z","timestamp":1542163361000},"page":"3580","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":30,"title":["Label-Free Quantitative Proteomics in a Methylmalonyl-CoA Mutase-Silenced Neuroblastoma Cell Line"],"prefix":"10.3390","volume":"19","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-6386-2009","authenticated-orcid":false,"given":"Michele","family":"Costanzo","sequence":"first","affiliation":[{"name":"Dipartimento di Medicina Molecolare e Biotecnologie Mediche, Universit\u00e0 degli Studi di Napoli \u201cFederico II\u201d, 80131 Naples, Italy"},{"name":"CEINGE\u2014Biotecnologie Avanzate s.c.ar.l., 80145 Naples, Italy"},{"name":"Associazione Culturale <i>DiSciMuS<\/i> RFC, Casoria, 80026 Naples, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1858-2833","authenticated-orcid":false,"given":"Armando","family":"Cevenini","sequence":"additional","affiliation":[{"name":"Dipartimento di Medicina Molecolare e Biotecnologie Mediche, Universit\u00e0 degli Studi di Napoli \u201cFederico II\u201d, 80131 Naples, Italy"},{"name":"CEINGE\u2014Biotecnologie Avanzate s.c.ar.l., 80145 Naples, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6960-8581","authenticated-orcid":false,"given":"Emanuela","family":"Marchese","sequence":"additional","affiliation":[{"name":"CEINGE\u2014Biotecnologie Avanzate s.c.ar.l., 80145 Naples, Italy"},{"name":"Dipartimento di Salute Mentale e Fisica e Medicina Preventiva, Universit\u00e0 degli Studi della Campania \u201cL. Vanvitelli\u201d, 80138 Naples, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Esther","family":"Imperlini","sequence":"additional","affiliation":[{"name":"IRCCS SDN, 80142 Naples, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Maddalena","family":"Raia","sequence":"additional","affiliation":[{"name":"CEINGE\u2014Biotecnologie Avanzate s.c.ar.l., 80145 Naples, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Luigi","family":"Del Vecchio","sequence":"additional","affiliation":[{"name":"CEINGE\u2014Biotecnologie Avanzate s.c.ar.l., 80145 Naples, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6035-8213","authenticated-orcid":false,"given":"Marianna","family":"Caterino","sequence":"additional","affiliation":[{"name":"Dipartimento di Medicina Molecolare e Biotecnologie Mediche, Universit\u00e0 degli Studi di Napoli \u201cFederico II\u201d, 80131 Naples, Italy"},{"name":"CEINGE\u2014Biotecnologie Avanzate s.c.ar.l., 80145 Naples, Italy"},{"name":"Associazione Culturale <i>DiSciMuS<\/i> RFC, Casoria, 80026 Naples, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7364-0602","authenticated-orcid":false,"given":"Margherita","family":"Ruoppolo","sequence":"additional","affiliation":[{"name":"Dipartimento di Medicina Molecolare e Biotecnologie Mediche, Universit\u00e0 degli Studi di Napoli \u201cFederico II\u201d, 80131 Naples, Italy"},{"name":"CEINGE\u2014Biotecnologie Avanzate s.c.ar.l., 80145 Naples, Italy"},{"name":"Associazione Culturale <i>DiSciMuS<\/i> RFC, Casoria, 80026 Naples, Italy"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2018,11,13]]},"reference":[{"key":"ref_1","unstructured":"Scriver, C.R., Beaudet, A.L., Sly, W.S., and Valle, D. 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