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Communication

Label-Free Quantitative Proteomics in a Methylmalonyl-CoA Mutase-Silenced Neuroblastoma Cell Line

1
Dipartimento di Medicina Molecolare e Biotecnologie Mediche, Università degli Studi di Napoli “Federico II”, 80131 Naples, Italy
2
CEINGE—Biotecnologie Avanzate s.c.ar.l., 80145 Naples, Italy
3
Associazione Culturale DiSciMuS RFC, Casoria, 80026 Naples, Italy
4
Dipartimento di Salute Mentale e Fisica e Medicina Preventiva, Università degli Studi della Campania “L. Vanvitelli”, 80138 Naples, Italy
5
IRCCS SDN, 80142 Naples, Italy
*
Authors to whom correspondence should be addressed.
Int. J. Mol. Sci. 2018, 19(11), 3580; https://doi.org/10.3390/ijms19113580
Received: 30 October 2018 / Accepted: 9 November 2018 / Published: 13 November 2018
(This article belongs to the Collection Advances in Proteomic Research)
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 β-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. View Full-Text
Keywords: quantitative proteomics; Methylmalonic Acidemias (MMAs); Methylmalonyl-CoA Mutase (MUT); energy metabolism; mitochondrial proteins quantitative proteomics; Methylmalonic Acidemias (MMAs); Methylmalonyl-CoA Mutase (MUT); energy metabolism; mitochondrial proteins
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MDPI and ACS Style

Costanzo, M.; Cevenini, A.; Marchese, E.; Imperlini, E.; Raia, M.; Del Vecchio, L.; Caterino, M.; Ruoppolo, M. Label-Free Quantitative Proteomics in a Methylmalonyl-CoA Mutase-Silenced Neuroblastoma Cell Line. Int. J. Mol. Sci. 2018, 19, 3580. https://doi.org/10.3390/ijms19113580

AMA Style

Costanzo M, Cevenini A, Marchese E, Imperlini E, Raia M, Del Vecchio L, Caterino M, Ruoppolo M. Label-Free Quantitative Proteomics in a Methylmalonyl-CoA Mutase-Silenced Neuroblastoma Cell Line. International Journal of Molecular Sciences. 2018; 19(11):3580. https://doi.org/10.3390/ijms19113580

Chicago/Turabian Style

Costanzo, Michele, Armando Cevenini, Emanuela Marchese, Esther Imperlini, Maddalena Raia, Luigi Del Vecchio, Marianna Caterino, and Margherita Ruoppolo. 2018. "Label-Free Quantitative Proteomics in a Methylmalonyl-CoA Mutase-Silenced Neuroblastoma Cell Line" International Journal of Molecular Sciences 19, no. 11: 3580. https://doi.org/10.3390/ijms19113580

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