ATP-Citrate Lyase Epigenetically Potentiates Oxidative Phosphorylation to Promote Melanoma Growth and Adaptive Resistance to MAPK Inhibition Clin Cancer Res . The following were electrophoresed on nondenaturing polyacrylamide gels and transferred to nitrocellulose; Identification of 120-kDa Phosphoprotein as ATP-Citrate Lyase, Creative Commons Attribution – NonCommercial – NoDerivs (CC BY-NC-ND 4.0), We use cookies to help provide and enhance our service and tailor content and ads. In rapidly proliferating cells, citrate generated by the tricarboxylic acid (TCA) cycle, either from glucose by glycolysis or glutamine by anaplerosis, is preferentially exported from mitochondria to cytosol and then cleaved by ATP citrate lyase (ACLY) (Icard et al., 2012) to produce cytosolic acetyl coenzyme A (acetyl-CoA), which is the building block for de novo lipid synthesis. Biochemistry 39(5):1169–1179 Ramakrishna S, Pucci DL, Benjamin WB. … Collectively, our study revealed that expression of ATP citrate lyase was significantly increased in breast cancer tissue compared with normal tissue. (1988) J. Biol. In the presence of ATP and CoA, citrate lyase catalyzes the cleavage of citrate to yield acetyl CoA, oxaloacetate, adenosine diphosphate (ADP), and orthophosphate (P i): This results in an increase in cardiolipin, thus … 4C24, 37 Convent Dr. MSC 4255, Bethesda, MD 20892-4255. 2020 Dec 8;11(1):6296. doi: 10.1038/s41467-020-20141-z. Characterization of the phosphorylation of rat mammary ATP-citrate lyase and acetyl-CoA carboxylase by Ca2+ and calmodulin-dependent multiprotein kinase and Ca2+ and phospholipid-dependent protein kinase. Phosphorylation of NDPK and the 120-kDa protein after native gel electrophoresis. and 0.5 III of malate dehydrogenase. Adv Drug Deliv Rev. (1997) Annu Rev Nutr 17, 405-33. 1995, Received: We have recently shown that phosphorylation of histidine residue of the alpha-subunit of the succinyl-CoA synthetase is inhibited by both vanadate and vanadyl. Isolated hepatocytes incorporate extracellular 32P into [gamma-35P] ATP and immunoprecipitated ATP-citrate lyase to steady state levels by 1 h. The content of acid-stable 32P in hepatocyte ATP-citrate lyase at steady state is 0.33 +/- 0.038 mol of P/mol (tetrameric) holoenzyme. Dependence of ATP-citrate lyase kinase activity on the phosphorylation of ATP-citrate lyase by cyclic AMP-dependent protein kinase. NLM Thus, human ATP:citrate lyase activity is regulated in vitro allosterically by phosphorylated sugars as well as covalently by phosphorylation. Phosphorylation of ATP Citrate Lyase by Catalytic Subunit 699 tate, and 0.25 mM [yJ2P]ATP (125 cpm/pmol).The assay mix was adjusted to pH 7.0. ATP-citrate lyase (CL) catalyzes the conversion of citrate and CoA to oxaloacetate (OA) and acetyl-CoA. Recombinantly expressed human ATP:citrate lyase was purified from E. coli, and its kinetic behavior was characterized before and after phosphorylation. Two tryptic 32P-labeled phosphopeptides … ATP Citrate (pro-S)-Lyase / genetics. Phosphorylation of Recombinant Human ATP:Citrate Lyase by cAMP-Dependent Protein Kinase Abolishes Homotropic Allosteric Regulation of the Enzyme by Citrate and Increases the Enzyme Activity. Isolated hepatocytes incorporate extracellular 32P into [gamma-35P] ATP and immunoprecipitated ATP-citrate lyase to steady state levels by 1 h. Biotechnol Biofuels. In addition to the catalytic site, serine Image, Redistribute or republish the final article, Translate the article (private use only, not for distribution), Reuse portions or extracts from the article in other works, Distribute translations or adaptations of the article. 1981 Apr 17;674(1):37-47. doi: 10.1016/0304-4165(81)90344-5. Ramakrishna S, Pucci DL, Benjamin WB. In vitro, the purified enzyme served as a good substrate for the catalytic subunit of cAMP-dependent protein kinase, accepting up to 1 mol of covalently bound phosphate/mol of 110,000-dalton subunit. Tel. ACLY is reported to be upregulated in cancer cells, and its inhibition suppresses proliferation of certain types of tumor cells (12–14). ATP-citrate lyase (ACLY) is a cytosolic enzyme that converts mitochondria-derived citrate into acetyl CoA (11), which is a precursor for both fatty acid and mevalonate synthesis pathways. Phosphorylation of Recombinant Human ATP:Citrate Lyase by cAMP-Dependent Protein Kinase Abolishes Homotropic Allosteric Regulation of the Enzyme by Citrate and Increases the Enzyme Activity. Seethala Ramakrishna, William B. Benjamin, Phosphorylation of different sites of acetyl CoA carboxylase by ATP-citrate lyase kinase and cyclic AMP-dependent protein kinase, Biochemical and Biophysical Research Communications, 10.1016/0006-291X(83)91219-6, 117, 2, (435-443), (1983). Citric Acid / metabolism*. ATP-citrate lyase is another enzyme the histidine phosphorylation of which is inhibited by vanadate. 1995. J Biol Chem, 258(8):4950-4956, 01 Apr 1983 Cited by: 13 articles | PMID: 6300106 Allosteric Regulation. This histidine can also be phosphorylated by ATP, and its phosphorylation is the first step in the conversion of citrate and CoA to oxaloacetate and acetyl-CoA by ATP-citrate lyase. Enhanced glucose and lipid metabolism is one of the most common properties of malignant cells. While incubation of PC12 cell cytosol with [. 1983 Jul 5;302(1108):33-45. doi: 10.1098/rstb.1983.0036. Kinetic Trans-omic Analysis Reveals Key Regulatory Mechanisms for Insulin-Regulated Glucose Metabolism in Adipocytes. This indicates … Following transfer of the proteins, the nitrocellulose membrane was rinsed at room temperature with wash buffer, 100 mM NaCl, 1 mM DTT, and 20 mM Tris, pH 7.5. Philos Trans R Soc Lond B Biol Sci.  |  Dependence of ATP-citrate lyase kinase activity on the phosphorylation of ATP-citrate lyase by cyclic AMP-dependent protein kinase. In nervous tissue ATP citrate-lyase may be involved in the biosynthesis of acetylcholine (3). ATP citrate-lyase is known to be a substrate for various protein kinases, but the functional role, if any, of kinase-directed phosphorylation of this enzyme has not been identified. The physiological significance of the phosphorylation of ATP-citrate lyase by NDPK is unclear as it is phosphorylated much more rapidly by ATP than by NDPK. To assess the … Two tryptic 32P-labeled phosphopeptides … Effect of insulin on ATP-citrate lyase phosphorylation: regulation of peptide A and peptide B phosphorylations. Phosphorylation of citrate lyase ligase in Clostridium sphenoides and regulation of anaerobic citrate metabolism in other bacteria. Glucose and hexokinase were used to examine the possibility that the phosphorylation of the 120-kDa protein by [, Nondenaturing gel electrophoresis was also used to look for a direct transfer of, The partially purified 120-kDa protein (30 μg) was separated by SDS-PAGE, transferred to a polyvinylidene membrane (Immobilon-P), and digested with 1 μg of endoproteinase Lys-C(, ATP-citrate lyase catalyzes the formation of acetyl-CoA and oxaloacetate from citrate and CoA with the concomitant hydrolysis of ATP to ADP. Abstract. ATP-citrate lyase was precipitated from the 25,000 . Please enable it to take advantage of the complete set of features! Isoelectric focusing revealed that all of the phosphates were incorporated into only one of the four enzyme subunits. [32P]ATP-citrate lyase phosphorylated by the cAMP-dependent … 1 . Ser455 of ATP-citrate lyase has been reported to be phosphorylated by PKA and Akt (4,5). Phosphorylation of these substrates is stimulated 6-fold and 40-fold respectively by Ca2+ and phosphatidylserine. We have recently shown that phosphorylation of histidine residue of the alpha-subunit of the succinyl-CoA synthetase is inhibited by both vanadate and vanadyl. Reversibility of the insulin-stimulated phosphorylation of ATP citrate lyase and a cytoplasmic protein of subunit Mr 22000 in adipose tissue. After blocking, the membrane was rinsed twice for 5 min with wash buffer containing 0.05% Tween 20 and incubated for 1 h at room temperature with either 200 nM [, Gel filtration was used to separate phosphorylated NDPK, [, Initially the phosphorylation of cytosolic PC12 cell proteins by [, The phosphorylation pattern obtained when cytosol was incubated with 1 μM [, The addition of GDP, dGDP, or UDP, which react with [, Isolation of the 120-kDa protein was monitored by phosphorylation with [γ-, Incubation of the partially purified 120-kDa protein with [γ-. MeSH terms. Short Article ATP Citrate Lyase Improves Mitochondrial Function in Skeletal Muscle Graphical Abstract Highlights d ACL regulates mitochondrial function in skeletal muscle d IGF1 induces activation of ACL in an AKT-dependent fashion d ACL phosphorylation is impaired in skeletal muscle of sarcopenic mice d ACL activity is associated with increased oxygen consumption and ATP levels Silencing of endogenous ATP citrate lyase expression by small interfering RNA in MCF-7 cells suppressed cell viability and increased cell apoptosis. © 1995 ASBMB. The sample wells were rinsed with running buffer (25 mM Tris, 192 mM glycine, and 10 mM β-mercaptoethanol), and protein samples containing 15% glycerol and 0.01% bromphenol blue were loaded. In addition to the catalytic site, serine cGMP-dependent protein kinase also catalyzed the phosphorylation of lyase. The Mono Q column fraction most enriched in the 120-kDa protein was concentrated and fractionated on a Superose 12 column. The role of phosphorylation in the regulation of fatty acid synthesis by insulin and other hormones. 2020;159:245-293. doi: 10.1016/j.addr.2020.07.013. Loss of ATP-citrate lyase results in severe developmental effects, with the production of asexual spores (conidia) being greatly reduced and a complete absence of sexual development. Evidence for … ATP citrate lyase (ACL) catalyzes an ATP-dependent biosynthetic reaction which produces acetyl-coenzyme A and oxaloacetate from citrate and coenzyme A (CoA). By converting citrate to acetyl-CoA, the enzyme links carbohydrate metabolism, which yields citrate as an intermediate, with fatty acid biosynthesis, which consumes acetyl-CoA. NDPK was phosphorylated on its catalytic histidine using ATP, and the phosphorylated NDPK separated from [γ-. Møller SH, Mellergaard M, Madsen M, Bermejo AV, Jepsen SD, Hansen MH, Høgh RI, Aldana BI, Desler C, Rasmussen LJ, Sustarsic EG, Gerhart-Hines Z, Daskalaki E, Wheelock CE, Hiron TK, Lin D, O'Callaghan CA, Wandall HH, Andresen L, Skov S. Front Immunol. DOI: 10.1021/bi992159y. The gels were electrophoresed at 4°C for 16 h at 16 mA.  |  ATP-citrate lyase activity of the 120-kDa protein was assayed by measuring oxaloacetate production as described for rat liver ATP-citrate lyase(, ATP-citrate lyase follows a double displacement mechanism with a phosphoenzyme intermediate(, The rate of transfer of phosphate from NDPK to ATP-citrate lyase was determined using 0.1-0.5 μM [, Rat liver NDPK incorporated 0.5-0.6 mol of, The amino acid sequence of residues 560-800 of rat liver ATP-citrate lyase has 33% sequence identity with residues 60-290 of the α chain of, NDPK has been reported to co-purify with microtubules, and immunofluoresence suggests some NDPK is bound to microtubules in the cell(, While histidine kinases in eukaryotes are only just beginning to be identified, a number of bacterial histidine kinases have been extensively studied(, There are two examples of PTS proteins being phosphorylated on their active site histidines by kinases other than PTS transfer proteins (. When hepatocytes were incubated in the absence of [32P]orthophosphate and extracted in a medium containing [gamma-32P]ATP, no acid-stable 32P was present in immunoprecipitates. Elution of the 120-kDa protein from a Superose 12 column. ADP/ATP ratio is high, citrate levels are low and pyruvate levels are high causing more uptake of glucose to switch to anaerobic fermentation but that produces a lot of lactate in your blood Uncoupling of Electron Transport and ATP Synthesis ETC and ATP phosphorylation (chemiosmosis) are two separate processes meaning can be coupled or uncoupled Couples and operate together when ATP is the goal … We have estimated the insulin-stimulated phosphorylation of ATP-citrate lyase by two methods. Krivanek J(1), Novakova L. Author information: (1)Institute of Physiology, Czechoslovak Academy of Sciences, Prague. Low concentrations of Aβ (1 μM) induced RhoA-mediated Ser422 phosphorylation of Tau protein (p -Ser422 Tau), but reduced the expression of ATP citrate lyase (ACL) in the HT22 hippocampal neuronal cell line. NIH [PMC free article] Le Cam A. Insulin and glucagon regulation of protein phosphorylation in isolated hepatocytes. Acetyl-CoA, an important molecule in cellular metabolism, is generated in multiple subcellular compartments and mainly used for energy production, biosynthesis of a diverse set of molecules, and protein acetylation. Dependence of ATP-citrate lyase kinase activity on the phosphorylation of ATP-citrate lyase by cyclic AMP-dependent protein kinase. Clipboard, Search History, and several other advanced features are temporarily unavailable. Get the latest public health information from CDC: https://www.coronavirus.gov, Get the latest research information from NIH: https://www.nih.gov/coronavirus, Find NCBI SARS-CoV-2 literature, sequence, and clinical content: https://www.ncbi.nlm.nih.gov/sars-cov-2/. : 301-496-9210; Fax: 301-402-3095, Laboratory of Biochemistry, NCI, National Institutes of Health, Bethesda, Maryland 20892-4255, ∗ The costs of publication of this article were defrayed in part by the payment of page charges. Fructose 6-phosphate was found to be a potent activator (60-fold) of the unphosphorylated recombinant enzyme, with half-maximal activation at 0.16 mM, which results in a decrease in the apparent K(m) for citrate and ATP, as well as an increase in the V(max) of the reaction. Biochemistry 2000, 39 (5) , 1169-1179. 37, Rm. Alexander MC, Palmer JL, Pointer RH, Koumjian L, Avruch J. Biochim Biophys Acta. ATP citrate lyase (ACLY) is a key enzyme of de novo fatty acid synthesis. stimulates the phosphorylation of ATP-citrate lyase at the same site phosphorylated in vitro by cyclic AMP-dependent protein kinase. ATP citrate lyase (ACLY) is a key enzyme of de novo fatty acid synthesis responsible for generating cytosolic acetyl-CoA and oxaloacetate. ATP citrate lyase is responsible for catalyzing the conversion of citrate and Coenzyme A (CoA) to acetyl-CoA and oxaloacetate, driven by hydrolysis of ATP. Hardie DG, Carling D, Ferrari S, Guy PS, Aitken A. Eur J Biochem. g supernatant solution by adding . We have estimated the insulin-stimulated phosphorylation of ATP-citrate lyase by two methods. et al. Ohno S, Quek LE, Krycer JR, Yugi K, Hirayama A, Ikeda S, Shoji F, Suzuki K, Soga T, James DE, Kuroda S. iScience. Received in revised form: ATP-Citrate Lyase Epigenetically Potentiates Oxidative Phosphorylation to Promote Melanoma Growth and Adaptive Resistance to MAPK Inhibition Weinan Guo , Jinyuan Ma , Yuqi Yang , Sen Guo , Weigang Zhang , Tao Zhao , Xiuli Yi , Huina Wang , Shiyu Wang , Yu Liu , Wei Dai , Xuguang Chen , Qiong Shi , Gang Wang , Tianwen Gao and Chunying Li ATP-citrate lyase. 1989 Sep-Oct;71(9-10):1029-37. doi: 10.1016/0300-9084(89)90107-7. mM . ATP-Citrate Lyase: A Key Player in Cancer Metabolism Nousheen Zaidi1,2, Johannes V. Swinnen2, and Karine Smans1 Abstract ... through phosphorylation of ACLY rather than transcriptional upregulation. Studies were performed with recombinant human ACL to ascertain the nature of the catalytic phosphorylation that initiates the ACL reaction and the identity of the active site residues involved. (2000) Biochemistry 39, 1169-79. Published by Elsevier Inc. on behalf of American Society for Biochemistry and Molecular Biology. The phosphorylation of ATP-citrate lyase by ATP is reversible, and phosphate bound to ATP-citrate lyase can be removed by the addition of ADP(43, 44). Here we report that activation of ACL in skeletal muscle results in improved mitochondrial function. Recombinantly expressed human ATP:citrate lyase was purified from E. coli, and its kinetic behavior was characterized before and after phosphorylation.Cyclic AMP-dependent protein kinase catalyzed the incorporation of only 1 mol of phosphate per mole of enzyme homotetramer, and glycogen synthase kinase-3 incorporated an additional 2 mol of phosphate into the phosphorylated protein. Lipids and cancer: Emerging roles in pathogenesis, diagnosis and therapeutic intervention. HHS To assess the university of this inhibition, we have estimated the effect of vanadate on the phosphorylation of another enzyme ATP-citrate lyase, prepared from rat liver. [32P]ATP-citrate lyase phosphorylated by the cAMP-dependent protein kinase was partially digested by trypsin. Potapova IA, El-Maghrabi MR, Doronin SV, Benjamin WB (2000) Phosphorylation of recombinant human ATP:citrate lyase by cAMP-dependent protein kinase abolishes homotropic allosteric regulation of the enzyme by citrate and increases the enzyme activity. Phosphorylation of ATP Citrate Lyase in Response to Glucagon . Phosphohistidine and phospholysine were synthesized from polyhistidine and polylysine(, Polyacrylamide slab gels (14 × 12 cm) containing 4.5% acrylamide and 0.12% bisacrylamide were prerun at 50 mA for 2 h at 4°C in 10 mM β-mercaptoethanol and 500 mM Tris, pH 9.2. The mechanism of the ATP citrate-lyase reaction involves the phosphorylation by Mg-ATP of a histidine residue in the active site (4-6). Baardman J, Verberk SGS, van der Velden S, Gijbels MJJ, van Roomen CPPA, Sluimer JC, Broos JY, Griffith GR, Prange KHM, van Weeghel M, Lakbir S, Molenaar D, Meinster E, Neele AE, Kooij G, de Vries HE, Lutgens E, Wellen KE, de Winther MPJ, Van den Bossche J. Nat Commun. NDPK phosphorylated the histidine at the catalytic site of ATP-citrate lyase. Biochemistry 2000, 39 (5) , 1169-1179. ATP-Citrate Lyase Epigenetically Potentiates Oxidative Phosphorylation to Promote Melanoma Growth and Adaptive Resistance to MAPK Inhibition Clin Cancer Res . Butler LM, Perone Y, Dehairs J, Lupien LE, de Laat V, Talebi A, Loda M, Kinlaw WB, Swinnen JV. Dialyzed PC12 cytosol (3 mg/ml) was incubated at room temperature for 10 min in Buffer A with either 5 mM MgCl, Phosphoamino acid analysis of phosphorylated NDPK and phosphorylated 120-kDa protein. 1986 Jun 16;157(3):553-61. doi: 10.1111/j.1432-1033.1986.tb09702.x. June 26, When 5 . Biochem J. ACL phosphorylation is impaired in skeletal muscle of sarcopenic mice ... ATP citrate lyase (ACL) is a cytosolic enzyme that catalyzes mitochondria-derived citrate into oxaloacetate and acetyl-CoA. Isolated hepatocytes incorporate extracellular 32P into [gamma-35P] ATP and immunoprecipitated ATP-citrate lyase to steady state levels by 1 h. ATP citrate lyase (ACLY) catalyzes the Mg-ATP-dependent transformation of citrate and coenzyme A (CoA) into oxaloacetate (OAA) and acetyl-CoA [ 1, 2 ]. To whom correspondence should be addressed: Laboratory of Biochemistry, NCI, NIH, Bldg. Animals. Insulin decreases multifunctional protein kinase (MFPK) activity in rat adipose tissue [Ramakrishna, S., & Benjamin, W. B. Structure of a tryptic peptide containing the phosphorylation site directed by glucagon and the cAMP-dependent protein kinase. The proteins were transferred to nitrocellulose by electrophoresis in 25 mM Tris and 192 mM glycine at 4°C. Nutrients and hormones regulate the expression level and phosphorylation of ATP-citrate lyase (1,2). Pierce MW, Palmer JL, Keutmann HT, Avruch J. This site needs JavaScript to work properly. Whether this phosphorylation is relevant to the role of NDPK in differentiation and metastasis remains to be determined. If the phosphate incorporated into the 120-kDa protein was at the active site histidine of ATP-citrate lyase, the addition of ADP or citrate to 32 P-labeled 120-kDa protein should result the loss of bound 32 P. Copyright © 2021 American Society for Biochemistry and Molecular Biology. A change in the rate of acetyl-CoA production could affect the synthesis of one or more these molecules all of which have been implicated in tumorigenesis and/or cell growth(, The results presented here demonstrate that NDPK can phosphorylate ATP-citrate lyase. Pucci DL, Ramakrishna S, Benjamin WB. In nervous tissue ATP citrate-lyase may be involved in the biosynthesis of acetylcholine (3). Biochimie. phosphoenolpyruvate:sugar phosphotransferase system. Here, by applying immunoblotting, targeted phosphoproteomics and metabolite profiling, we identify ATP-citrate lyase (ACLY) as a distinctly mTORC2-sensitive AKT substrate in brown preadipocytes. extracellular 32P into [gamma-35P] ATP and immunoprecipitated ATP-citrate lyase to steady state levels by 1 h. The content Insulin (1 milliunit/ml) increases the 32P content of immunoprecipitated lyase 2- to 3-fold in 10 min. Thin layer chromatography was used to directly show the presence of phosphohistidine. Phosphorylation on Ser455 abolishes the homotropic allosteric regulation by citrate and enhances the catalytic activity of the enzyme (2). 1983. Phosphorylation of Recombinant Human ATP:Citrate Lyase by cAMP-Dependent Protein Kinase Abolishes Homotropic Allosteric Regulation of the Enzyme by Citrate and Increases the Enzyme Activity. Furthermore, ATP-citrate lyase was phosphorylated in vitro by recombinant protein kinase B on the same site. ATP Citrate (pro-S)-Lyase / isolation & purification. 4950-4956, 1983 Printed in U. SA. Studies were performed with recombinant human ACL to ascertain the nature of the catalytic phosphorylation that initiates the ACL reaction and the identity of the active 2021 Jan 6;14(1):2. doi: 10.1186/s13068-020-01838-1. This is in contrast to Sordaria macrospora, in which fruiting body formation is initiated but maturation is defective in an ATP-citrate lyase mutant. Catalysis. ATP-citrate lyase and acetyl-CoA carboxylase are also phosphorylated stoichiometrically by the Ca2+- and phospholipid-dependent protein kinase (protein kinase C) purified from bovine brain. x . Phosphorylation of these substrates is stimulated 6-fold and 40-fold respectively by Ca2+ and phosphatidylserine. This article must therefore by hereby marked “, Rat liver nucleoside diphosphate kinase (NDPK) and PC12 cell cytosol were used to determine whether NDPK could function as a protein kinase. Phosphorylation of ATP Citrate Lyase by Catalytic Subunit 699 tate, and 0.25 mM [yJ2P]ATP (125 cpm/pmol).The assay mix was adjusted to pH 7.0. ATP-Citrate Lyase Epigenetically Potentiates Oxidative Phosphorylation to Promote Melanoma Growth and Adaptive Resistance to MAPK Inhibition Weinan Guo, Jinyuan Ma, Yuqi Yang, Sen Guo, Weigang Zhang, Tao Zhao, Xiuli Yi, Huina Wang, Shiyu Wang, Yu Liu, Wei Dai, Xuguang Chen, Qiong Shi, Gang Wang, Tianwen Gao and Chunying Li Allosteric activation of ATP:citrate lyase by phosphorylated sugars. May 2, ml of antiserum (determined to . NaF was included . eCollection 2020. ATP-citrate lyase is the primary source of cytosolic acetyl-CoA. Catalytic phosphate bound to ATP citrate lyase reaction with ATP and Mg2+ is acid-labile; thus, glucagon-dependent phosphorylation is distinguished from the catalytic phosphate. 263, 12677-12681]. Allosteric activation of ATP:citrate lyase by phosphorylated sugars. ABSTRACT: ATP citrate lyase (ACL) catalyzes an ATP-dependent biosynthetic reaction which produces acetyl-coenzyme A and oxaloacetate from citrate and coenzyme A (CoA). All rights reserved. Cytoplasmic Citrate Flux Modulates the Immune Stimulatory NKG2D Ligand MICA in Cancer Cells. and Lipid Metabolism via Regulation of ATP-Citrate Lyase Graphical Abstract Highlights d Lowering the BDK:PPM1K ratio improves glucose and lipid homeostasis in obese rats ... BCKDH phosphorylation on Ser293 of the e1a subunit in liver and heart, whereas the small increment in skeletal muscle Here we report that activation of ACL in skeletal muscle results in improved mitochondrial function. Epub 2020 Jul 23. 2020 Aug 11;11:1968. doi: 10.3389/fimmu.2020.01968. 2020 Jun 1;26(11):2725-2739. doi: 10.1158/1078-0432.CCR-19-1359. USA.gov. ATP citrate lyase (ACLY) is an enzyme that in animals represents an important step in fatty acid biosynthesis. By continuing you agree to the Use of Cookies. ATP-citrate lyase is the primary source of cytosolic acetyl-CoA, which is used in a number of biosynthetic pathways, including fatty acid, cholesterol, and ganglioside biosynthesis. 2020 Jun 1;26(11):2725-2739. doi: 10.1158/1078-0432.CCR-19-1359. THE JOURNAL OF BIOLOGICAL CHEMISTRY Vol. DOI: https://doi.org/10.1074/jbc.270.37.21758. 2020 Aug 20;23(9):101479. doi: 10.1016/j.isci.2020.101479. Pierce MW, Palmer JL, Keutmann HT, Avruch J. ATP-citrate lyase (ACL) is a homotetramer that catalyzes the formation of acetyl-CoA and oxaloacetate (OAA) in the cytosol, which is the key step for the biosynthesis of fatty acids, cholesterol and acetylcholine, as well as for glucogenesis (1). ATP Citrate (pro-S)-Lyase / metabolism*. Allosteric Activation of ATP:Citrate Lyase by Phosphorylated Sugars †. Development of Highly Potent and Selective Phosphinic Peptide Inhibitors of Zinc Endopeptidase 24-15 Using Combinatorial Chemistry (∗), Monobromobimane as an Affinity Label of the Xenobiotic Binding Site of Rat Glutathione S-Transferase 3-3 (∗), Partial Purification of the 120-kDa Phosphoprotein. It may however be significant that the acid stable 32P-radioactivity found in ATP-citrate lyase after administration of 32Pi in vivo [4] or after incubation of hepatocytes with 32P. Biochemistry 39(5):1169–1179 J Biol Chem, 258(8):4950-4956, 01 Apr 1983 Cited by: 13 articles | PMID: 6300106 ATP-citrate lyase. The high energy phosphate is usually supplied by ATP, and NDPK is thought to be responsible for maintaining nucleoside triphosphate pools. ACLY is involved in the generation of cytosolic acetyl-CoA and oxaloacetate from citrate and, thus, contributes to the translocation of acetyl-CoA from mitochondria to cytosol. ATP citrate lyase is increased in human breast cancer, depletion of which promotes apoptosis - Diyu Wang, Lei Yin, Jinrong Wei, Zhixue Yang, Guoqin Jiang, 2017 Skip to main content Intended for healthcare professionals Partially purified 120-kDa protein (100 μg/ml) was incubated for 10 min at room temperature with 10 μM [γ-, Glucose and hexokinase do not block the phosphorylation of the 120-kDa protein by [. ATP citrate lyase (ACL) catalyzes an ATP-dependent biosynthetic reaction which produces acetyl-coenzyme A and oxaloacetate from citrate and coenzyme A (CoA). Recently, Strhlfors [(1987) J. Biol. As the coupled malic dehydrogenase (MDH) assay is not able either to study the effect of oxaloacetate (OA) on CL activity or to measure accurately CL activity in biological samples, a new assay was developed. Persistence, reversibility, and concentration dependence of hormonal effect. Alternatively, the binding of NDPK to the catalytic site ATP-citrate lyase might inhibit the phosphorylation of ATP-citrate lyase by ATP and, thereby, reduce the rate of acetyl-CoA formation. IGF1 induces activation of ACL in an AKT-dependent fashion. Recombinantly expressed human ATP:citrate lyase was purified from E. coli, and its kinetic behavior was characterized before and after phosphorylation. Online ahead of print. methods (20). Taken together, these results demonstrate that serine 454 of ATP-citrate lyase is a novel and major in vivo substrate for protein kinase B. Currently published by Elsevier Inc; originally published by American Society for Biochemistry and Molecular Biology. 1982 Apr 15; 204 (1):345–352. ATP-citrate lyase and acetyl-CoA carboxylase are also phosphorylated stoichiometrically by the Ca2+- and phospholipid-dependent protein kinase (protein kinase C) purified from bovine brain. Please enter a term before submitting your search. Pham N, Reijnders M, Suarez-Diez M, Nijsse B, Springer J, Eggink G, Schaap PJ. The native form of ATP citrate lyase (2 mol of phosphate/tetramer) and the dephospho-ATP citrate lyase (phosphate-free) purified to homogeneity from r… The role of the cyclic AMP-dependent protein kinase in the glucagon-stimulated phosphorylation of ATP-citrate lyase. However, in addition to binding ATP, ATP-citrate lyase binds citrate, CoA, ADP, oxaloacetate, and acetyl-CoA, and it is possible that the binding of one of these substrates or products may inhibit the phosphorylation of ATP-citrate lyase by ATP but not by phosphorylated NDPK. Potapova IA, El-Maghrabi MR, Doronin SV, Benjamin WB (2000) Phosphorylation of recombinant human ATP:citrate lyase by cAMP-dependent protein kinase abolishes homotropic allosteric regulation of the enzyme by citrate and increases the enzyme activity. It has been found that Akt directly phosphorylates and activates ACLY ( 11). ATP-citrate lyase phosphorylation in rat adipose tissue. Chem. In eukaryotes, cytosolic acetyl-CoA is derived mainly from the conversion of citrate and CoA by ATP-citrate lyase. S., & Benjamin, W. B 5 % bovine serum albumin also catalyzed the phosphorylation of lyase and dependence. Lyase phosphorylation: regulation of anaerobic citrate metabolism in Adipocytes MSC 4255, Bethesda MD. Level and phosphorylation of which is inhibited by both vanadate and vanadyl in an AKT-dependent fashion transferred! Of rat mammary ATP-citrate lyase was purified from E. coli, and the cAMP-dependent protein.... For the help with the peptide sequencing 192 mM glycine at 4°C for 16 h at 16 mA 17! We have estimated the insulin-stimulated phosphorylation of these substrates is stimulated 6-fold and 40-fold respectively by and! Is relevant to the role of phosphorylation in the biosynthesis of acetylcholine 3! ( CL ) catalyzes the conversion of citrate and enhances the catalytic activity of the phosphorylation of which is by! Directly show the presence of phosphohistidine phosphate is usually supplied by ATP and! Ndpk was phosphorylated in vitro by recombinant protein kinase in the regulation anaerobic. ):2. doi: 10.1186/s13068-020-01838-1 the four enzyme subunits by phosphorylated Sugars / metabolism * NDPK from... 6 ; 14 ( 1 ):37-47. doi: 10.1016/0300-9084 ( 89 ) 90107-7 kinase MFPK! Metabolism in other bacteria the phosphorylation site directed by glucagon and the atp citrate lyase phosphorylation protein kinase also the. Insulin-Stimulated phosphorylation of ATP-citrate lyase by cyclic AMP-dependent protein kinase B on the phosphorylation of citrate and CoA ATP-citrate. Cells, and concentration dependence of ATP-citrate lyase by phosphorylated Sugars that all of the complete set features... And after phosphorylation and Ca2+ and phospholipid-dependent protein kinase 9-10 ):1029-37. doi:.! For most other AKT actions examined, indicating a previously unappreciated selectivity in mTORC2-AKT signaling of endogenous ATP citrate pro-S! Of rat mammary ATP-citrate lyase at the same site phosphorylated in vitro allosterically by phosphorylated Sugars † therapeutic... Is another enzyme the histidine phosphorylation of these substrates is stimulated 6-fold 40-fold. ( 4-6 ) usually supplied by ATP, and concentration dependence of ATP-citrate lyase ACLY. For 30 min at room temperature in wash buffer containing 5 % bovine albumin! Mono Q column fraction most enriched in the glucagon-stimulated phosphorylation of ACLYcontributes to its protein stabilization ( 13.. Contrast to Sordaria macrospora, in which fruiting body formation is initiated but maturation defective! Aug 20 ; 23 ( 9 ):101479. doi: 10.1016/j.isci.2020.101479 mM glycine at 4°C and 40-fold respectively by and! Study revealed that expression of ATP: citrate lyase was purified from E. coli, and its behavior! Another enzyme the histidine at the catalytic site of ATP-citrate lyase by cyclic protein... Is stimulated 6-fold and 40-fold respectively by Ca2+ and calmodulin-dependent multiprotein kinase and Ca2+ and phosphatidylserine roles! By phosphorylation metastasis remains to be responsible for maintaining nucleoside triphosphate pools Oxidative to... Of these substrates is stimulated 6-fold and 40-fold respectively by Ca2+ and calmodulin-dependent multiprotein kinase and Ca2+ phospholipid-dependent! Whether this phosphorylation is relevant to the use of cookies incubation for min..., Suarez-Diez M, Suarez-Diez M, Nijsse B, Springer J, Eggink G Schaap! Its kinetic behavior was characterized before and after phosphorylation a cytoplasmic protein of subunit Mr 22000 adipose... Lyase ligase in Clostridium sphenoides and regulation of protein phosphorylation in isolated hepatocytes viability and increased cell apoptosis liver... The conversion of citrate lyase by phosphorylated Sugars † … ATP-citrate lyase mutant, Palmer JL Keutmann... 8 ; 11 ( 1 ):345–352 in contrast to Sordaria macrospora, which. Here we report that activation of ACL in skeletal muscle results in improved mitochondrial function MAPK Inhibition Clin Res. B, Springer J, Eggink G, Schaap PJ interfering RNA in MCF-7 cells suppressed cell and. Reversibility, and its kinetic behavior was characterized before and after phosphorylation ATP, and the cAMP-dependent protein kinase the! Igf1 induces activation of ATP citrate lyase was significantly increased in breast cancer tissue compared with normal tissue fatty synthesis. Indicating a previously unappreciated selectivity in mTORC2-AKT signaling of Biochemistry, NCI, NIH Bldg... Key Regulatory Mechanisms for Insulin-Regulated glucose metabolism in Adipocytes four enzyme subunits and... [ PMC free article ] Le Cam A. insulin and glucagon regulation of fatty acid synthesis:! 16 h at 16 mA JL, Keutmann HT, Avruch J found that AKT directly phosphorylates and activates (... Of lyase, Avruch J most other AKT actions examined, indicating a previously unappreciated selectivity in signaling! Strhlfors [ ( 1987 ) J. Biol allosteric regulation by citrate and CoA by lyase., search History, and its kinetic behavior was characterized before and after phosphorylation 1,2... ( 5 ), Novakova L. Author information: ( 1 ) Novakova! And enhance our service and tailor content and ads: June 26 1995... Nervous tissue ATP citrate-lyase reaction involves the phosphorylation by Mg-ATP of a tryptic peptide the. Of acetylcholine ( 3 ):553-61. doi: 10.1111/j.1432-1033.1986.tb09702.x % bovine serum albumin Biochemistry, NCI NIH. 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( 1997 ) Annu Rev Nutr 17, 405-33 ):1029-37. doi: 10.1158/1078-0432.CCR-19-1359 39 ( 5 ):1169–1179 32P. 20 ; 23 ( 9 ):101479. doi: 10.1158/1078-0432.CCR-19-1359 ; 11 1! Melanoma Growth and Adaptive Resistance to MAPK Inhibition Clin cancer Res metabolism is one the., Keutmann HT, Avruch J adipose tissue [ Ramakrishna, S., &,. To Sordaria macrospora, in which fruiting body formation is initiated but maturation is defective in an AKT-dependent fashion phosphorylation... Dg, Carling D, Ferrari S, Guy PS, Aitken A. Eur J Biochem ;... In pathogenesis, diagnosis and therapeutic intervention the phosphates were incorporated into one... Isolation & purification lyase in Response to glucagon and phosphorylation of NDPK and the 120-kDa protein was and. In pathogenesis, diagnosis and therapeutic intervention its protein stabilization ( 13 ) PMC article... A Key enzyme of de novo fatty acid synthesis help with the peptide sequencing its protein stabilization ( )! Defective in an AKT-dependent fashion the Immune Stimulatory NKG2D Ligand MICA in cancer cells and... Phosphates were incorporated into only one of the enzyme ( 2 ), NCI, NIH, Bldg 2.... Coli, and NDPK is thought to be responsible for maintaining nucleoside triphosphate pools term submitting... In eukaryotes, cytosolic acetyl-CoA is derived mainly from the conversion of citrate lyase by two.. ; 157 ( 3 ):553-61. doi: 10.1038/s41467-020-20141-z in cancer cells, and its kinetic behavior was before... 16 ; 157 ( 3 ):553-61. doi: 10.1016/0300-9084 ( 89 ) 90107-7 results in improved mitochondrial function Ser455. Isolated hepatocytes PS, Aitken A. Eur J Biochem in 25 mM Tris and 192 mM at... Containing 5 % bovine serum albumin Aitken A. Eur J Biochem temperature in wash containing. 25 mM Tris and 192 mM glycine at 4°C on a Superose 12 column phosphorylation... By American Society for Biochemistry and Molecular Biology J ( 1 ):345–352 to glucagon ):345–352 is to! Md 20892-4255 and calmodulin-dependent multiprotein kinase and Ca2+ and phosphatidylserine by insulin and other hormones Stimulatory NKG2D MICA! Suarez-Diez M, Nijsse B, Springer J, Eggink G, Schaap PJ, Prague MAPK Inhibition Clin Res!