PMID- 37224468 OWN - NLM STAT- MEDLINE DCOM- 20230630 LR - 20231014 IS - 1522-1555 (Electronic) IS - 0193-1849 (Print) IS - 0193-1849 (Linking) VI - 325 IP - 1 DP - 2023 Jul 1 TI - A high-fat diet increases hepatic mitochondrial turnover through restricted acetylation in a NAFLD mouse model. PG - E83-E98 LID - 10.1152/ajpendo.00310.2022 [doi] AB - Lysine acetylation of proteins has emerged as a key posttranslational modification (PTM) that regulates mitochondrial metabolism. Acetylation may regulate energy metabolism by inhibiting and affecting the stability of metabolic enzymes and oxidative phosphorylation (OxPhos) subunits. Although protein turnover can be easily measured, due to the low abundance of modified proteins, it has been difficult to evaluate the effect of acetylation on the stability of proteins in vivo. We applied (2)H(2)O-metabolic labeling coupled with immunoaffinity and high-resolution mass spectrometry method to measure the stability of acetylated proteins in mouse liver based on their turnover rates. As a proof-of-concept, we assessed the consequence of high-fat diet (HFD)-induced altered acetylation in protein turnover in LDL receptor-deficient (LDLR(-/-)) mice susceptible to diet-induced nonalcoholic fatty liver disease (NAFLD). HFD feeding for 12 wk led to steatosis, the early stage of NAFLD. A significant reduction in acetylation of hepatic proteins was observed in NAFLD mice, based on immunoblot analysis and label-free quantification with mass spectrometry. Compared with control mice on a normal diet, NAFLD mice had overall increased turnover rates of hepatic proteins, including mitochondrial metabolic enzymes (0.159 +/- 0.079 vs. 0.132 +/- 0.068 day(-1)), suggesting their reduced stability. Also, acetylated proteins had slower turnover rates (increased stability) than native proteins in both groups (0.096 +/- 0.056 vs. 0.170 +/- 0.059 day(-1) in control, and 0.111 +/- 0.050 vs. 0.208 +/- 0.074 day(-1) in NAFLD). Furthermore, association analysis revealed a relationship between the HFD-induced decrease in acetylation and increased turnover rates for hepatic proteins in NAFLD mice. These changes were associated with increased expressions of the hepatic mitochondrial transcriptional factor (TFAM) and complex II subunit without any changes to other OxPhos proteins, suggesting that enhanced mitochondrial biogenesis prevented restricted acetylation-mediated depletion of mitochondrial proteins. We conclude that decreased acetylation of mitochondrial proteins may contribute to adaptive improved hepatic mitochondrial function in the early stages of NAFLD.NEW & NOTEWORTHY This is the first method to quantify acetylome dynamics in vivo. This method revealed acetylation-mediated altered hepatic mitochondrial protein turnover in response to a high-fat diet in a mouse model of NAFLD. FAU - Aghayev, Mirjavid AU - Aghayev M AD - Department of Pharmaceutical Sciences, College of Pharmacy, Northeast Ohio Medical University, Rootstown, Ohio, United States. FAU - Arias-Alvarado, Andrea AU - Arias-Alvarado A AUID- ORCID: 0000-0003-1362-443X AD - Department of Pharmaceutical Sciences, College of Pharmacy, Northeast Ohio Medical University, Rootstown, Ohio, United States. FAU - Ilchenko, Sergei AU - Ilchenko S AD - Department of Pharmaceutical Sciences, College of Pharmacy, Northeast Ohio Medical University, Rootstown, Ohio, United States. FAU - Lepp, Josephine AU - Lepp J AD - Department of Pharmaceutical Sciences, College of Pharmacy, Northeast Ohio Medical University, Rootstown, Ohio, United States. FAU - Scott, Iain AU - Scott I AUID- ORCID: 0000-0001-5929-0928 AD - Cardiology Division, Department of Medicine, University of Pittsburgh Medical Center, Pittsburgh, Pennsylvania, United States. FAU - Chen, Yeong-Renn AU - Chen YR AUID- ORCID: 0000-0001-5511-8215 AD - Department of Integrative Medical Sciences, College of Medicine, Northeast Ohio Medical University, Rootstown, Ohio, United States. FAU - Zhang, Guo-Fang AU - Zhang GF AUID- ORCID: 0000-0003-3484-5864 AD - Division of Endocrinology, Metabolism and Nutrition, Duke Molecular Physiology Institute, Duke University, Durham North Carolina, United States. AD - Department of Medicine, Duke University, Durham North Carolina, United States. FAU - Tsai, Tsung-Heng AU - Tsai TH AD - Department of Mathematical Sciences, Kent State University, Kent, Ohio, United States. FAU - Kasumov, Takhar AU - Kasumov T AUID- ORCID: 0000-0003-2173-7231 AD - Department of Pharmaceutical Sciences, College of Pharmacy, Northeast Ohio Medical University, Rootstown, Ohio, United States. LA - eng SI - figshare/10.6084/m9.figshare.21606513 GR - R01 HL147861/HL/NHLBI NIH HHS/United States GR - R01 HL129120/HL/NHLBI NIH HHS/United States GR - R01 GM112044/GM/NIGMS NIH HHS/United States GR - R01 AA030026/AA/NIAAA NIH HHS/United States GR - R01 HL156874/HL/NHLBI NIH HHS/United States GR - R21 AA029784/AA/NIAAA NIH HHS/United States PT - Journal Article PT - Research Support, N.I.H., Extramural DEP - 20230524 PL - United States TA - Am J Physiol Endocrinol Metab JT - American journal of physiology. Endocrinology and metabolism JID - 100901226 RN - 0 (Mitochondrial Proteins) SB - IM MH - Animals MH - Mice MH - *Non-alcoholic Fatty Liver Disease/metabolism MH - Diet, High-Fat MH - Acetylation MH - Liver/metabolism MH - Protein Processing, Post-Translational MH - Mitochondrial Proteins/metabolism MH - Mitochondrial Turnover MH - Mice, Inbred C57BL PMC - PMC10312330 OTO - NOTNLM OT - NAFLD OT - acetylome dynamics OT - metabolic labeling OT - mitochondria OT - turnover COIS- No conflicts of interest, financial or otherwise, are declared by the authors. EDAT- 2023/05/24 19:13 MHDA- 2023/06/30 06:42 PMCR- 2024/07/01 CRDT- 2023/05/24 16:42 PHST- 2024/07/01 00:00 [pmc-release] PHST- 2023/06/30 06:42 [medline] PHST- 2023/05/24 19:13 [pubmed] PHST- 2023/05/24 16:42 [entrez] AID - E-00310-2022 [pii] AID - 10.1152/ajpendo.00310.2022 [doi] PST - ppublish SO - Am J Physiol Endocrinol Metab. 2023 Jul 1;325(1):E83-E98. doi: 10.1152/ajpendo.00310.2022. Epub 2023 May 24.