Language links are at the top of the page across from the title. Succinyl-CoA ligase [ADP-forming] subunit beta, mitochondrial (SUCLA2), also known as ADP-forming succinyl-CoA synthetase (SCS-A), is an enzyme that in humans is encoded by the SUCLA2 gene on chromosome 13. We therefore evaluated NTP synthesis by the 16- and 12-kDa forms of Ndk in the presence of the and subunits of Scs. Two forms of Ndk, a 16-kDa form and a truncated 12-kDa form obtained by treatment with purified elastase (9) were used. In eukaryotes, the Scs occurs predominantly in the G-form, where it is a heterodimer composed of the and subunits (8, 25). The enzyme from Escherichia coli is an alpha 2 beta 2 tetramer (mol. Succinyl Coenzyme A Synthetase - an overview - ScienceDirect Inclusion in an NLM database does not imply endorsement of, or agreement with, Succinyl-CoA ligase, also called succinate synthase, is an enzyme in the Krebs cycle that converts succinyl-CoA to succinate and free CoA, and converts ADP or guanosine diphosphate (GDP) to ATP or . Succinyl-Coenzyme A to Oxaloacetate Succinate thiokinase. Similarly, 5 M ADP in the reaction mixture weakly enhanced the phosphorylation, while 50 M ADP did not have any effect (data not shown). The crystal structure of Succinyl-CoA synthetase alpha subunit (succinyl-CoA-binding isoform) was determined by Joyce et al. Escherichia coli succinyl coenzyme A synthetase. Surendran S, Adaikalakoteswari A, Saravanan P, Shatwaan IA, Lovegrove JA, Vimaleswaran KS. A 20-kb EcoRI cosmid clone (pSU6) from P. aeruginosa PAO1 genomic library that hybridized to a 512-bp sucCD-specific probe (14) was isolated. Simulation results of mechanism D, partial random sequential kinetic mechanism with enzyme-bound succinyl, Figure A6. The specificity for either adenosine or guanosine phosphates is defined by the -subunit, [10] which is encoded by 2 genes. Kamath S, Kapatral V, Chakrabarty A M. Cellular function of elastase in, Kavanaugh-Black A, Connolly D M, Chugani S A, Chakrabarty A M. Characterization of nucleoside diphosphate kinase from. Based on these results, we conclude that the Scs enzyme exhibits NDP kinase activity with GDP = ADP > UDP > CDP with respect to its relative affinities for the NDPs. An official website of the United States government. [17], Measurements (performed using a soy bean SCS) indicate an optimal temperature of 37C and an optimal pH of 7.0-8.0. Edmonton, AB, Canada: University of Alberta; 2000. Succinyl-coenzyme A, abbreviated as succinyl-CoA or SucCoA, is a thioester of succinic acid and coenzyme A. Fatal infantile lactic acidosis: Defective SCS has been implicated as a cause of fatal infantile lactic acidosis, which is a disease in infants that is characterized by the build-up of toxic levels of lactic acid. [3] The enzyme facilitates the coupling of this reaction to the formation of a nucleoside triphosphate molecule from an inorganic phosphate molecule and a nucleoside diphosphate molecule . In biochemistry, a ligase is an enzyme that can catalyze the joining of two molecules by forming a new chemical bond.This is typically via hydrolysis of a small pendant chemical group on one of the molecules, typically resulting in the formation of new C-O, C-S, or C-N bonds. A pertinent example is the synthesis of the polysaccharide alginate by P. aeruginosa cells isolated from the lungs of CF patients (16). -, Alarcon C., Wicksteed B., Prentki M., Corkey B. E., Rhodes C. J. Succinate is a preferential metabolic stimulus-coupling signal for glucose-induced proinsulin biosynthesis translation. Clipboard, Search History, and several other advanced features are temporarily unavailable. succinyl coenzyme A synthetase preparation from Esch- erichiacoli has the following characteristics:apparent by incubation with CoA, and the occurrence of a Pi-independent succinate F) succinyl-CoA exchange led to suggestion of t,he followingreaction sequence where E designatesfree enzyme eCollection 2018. [9] The formation of the nucleotide triphosphate occurs in an ATP grasp domain, which is located near the N-terminus of the each subunit. In those experiments, [18O]P Even though it is branded as a 'cycle', it is not necessary for metabolites to follow only one specific route; at least three alternative segments of the citric acid cycle have been recognized. The role of succinyl-CoA synthetase in the control of heme biosynthesis. However, both ATP- and GTP-specific Scs isoforms have been characterized from multicellular eukaryotes (8) and, in general, the nucleotide preference varies widely within organisms (11, 20). The presence of Scs , , or both in association with the 16-kDa Ndk did not lead to any alteration of NTP synthesis (Fig. SUCLG2 is GTP-specific and SUCLA2 is ATP-specific, while SUCLG1 encodes the common -subunit. A loss of phosphorylation of the subunit is an indication of the binding of the nucleotide to the catalytic site of the -subunit of Scs. Pall M L. GTP, a central regulator of cellular anabolism. Such a mechanism of GTP synthesis can be an alternate source for the synthesis of alginate as well as for the synthesis of other macromolecules requiring GTP such as RNA and protein. 2018 Feb 6;13:2. doi: 10.1186/s12263-018-0591-9. 8600 Rockville Pike Succinyl Coenzyme A - an overview | ScienceDirect Topics Succinyl Coenzyme A Add to Mendeley Set alert Nitric Oxide, Part G Oxidative and Nitrosative Stress in Redox Regulation of Cell Signaling Catherine Bregere, . Accessibility Epub 2021 May 10. Before The Scs subunit can be efficiently autophosphorylated either in presence of ATP or in presence of GTP, and small quantities of GDP or ADP can enhance such phosphorylation, while larger concentrations inhibit it, suggesting that intracellular concentrations of ADP and GDP modulate the interconversion of ATP and GTP by Scs. [17], Measurements (performed using a soy bean SCS) indicate an optimal temperature of 37C and an optimal pH of 7.0-8.0.[18]. studies) suggest that co-operative interactions between alternatingly functional active sites on the two halves of the E. coli enzyme contribute to its catalytic efficacy. The citric acid cycle (CAC)also known as the Krebs cycle, Szent-Gyrgyi-Krebs cycle or the TCA cycle (tricarboxylic acid cycle)is a series of chemical reactions to release stored energy through the oxidation of acetyl-CoA derived from carbohydrates, fats, and proteins. Identification of the kinetic mechanism of succinyl-CoA synthetase Succinyl-CoA ligase [GDP-forming] subunit beta, mitochondrial is an enzyme that in humans is encoded by the SUCLG2 gene on chromosome 3. PDB, "Interactions of GTP with the ATP-grasp domain of GTP-specific succinyl-CoA synthetase", "Genetic evidence for the expression of ATP- and GTP-specific succinyl-CoA synthetases in multicellular eucaryotes", "Expression of two succinyl-CoA synthetases with different nucleotide specificities in mammalian tissues", "Characterization of the ATP- and GTP-specific succinyl-CoA synthetases in pigeon. The reaction catalyzed by succinyl-CoA synthetase (E.C. Both synthesis and degradation reactions require multiple enzymes to facilitate the event. Nishimura J S. Succinyl-CoA synthetase structure-function relationships and other considerations. Succinyl-CoA synthetase structure-function relationships and other considerations. As shown in Fig. [3] The enzyme facilitates the coupling of this reaction to the formation of a nucleoside triphosphate molecule (either GTP or ATP) from an inorganic phosphate molecule and a nucleoside diphosphate molecule (either GDP or ADP). Liu M, Han X, Xian M, Ding Y, Liu H, Zhao G. Bioengineered. Simulation results of mechanism A, partial random sequential kinetic mechanism with enzyme-bound phosphoryl, Figure 4. To further delineate the relative affinity of ADP and GDP as substrates, a substrate competition experiment was conducted at various concentrations in the presence of inorganic [32P]Pi, succinyl-CoA, and GDP or ADP. HHS Vulnerability Disclosure, Help Succinyl Coenzyme A - an overview | ScienceDirect Topics A 5 M concentration of GDP in the reaction mixture stimulated the phosphorylation of the 30-kDa Scs subunit, whereas 50 M GDP in the reaction mixture showed a significant decrease. Since the intracellular level of NDPs may reach millimolar concentrations, it is conceivable that P. aeruginosa Scs may generate all of the NTPs under certain conditions. Finally, the phosphorylated histidine transfers the phosphate group to a nucleoside diphosphate, which generates the high-energy carrying nucleoside triphosphate. and transmitted securely. The general reaction via ping-pong mechanism is as follows: XDP + YTP XTP + YDP. The first enzyme, succinyl coenzyme A (succinyl-CoA):acetate CoA-transferase (SCACT), forms acetate from acetyl-CoA. By means of active enzyme centrifugation, we have shown that the active form of the bacterial enzyme is the tetramer even at very low assay concentrations, while the pig heart enzyme is a non-associating dimer over a wide concentration range. The purified Scs subunits were incubated with various amounts of succinate and CoA () or with succinate only () using [-32P]ATP as a phosphodonor. The strains and plasmids used in this study are listed in Table Table1.1. Adenosine triphosphate (ATP) is an organic compound that provides energy to drive and support many processes in living cells, such as muscle contraction, nerve impulse propagation, condensate dissolution, and chemical synthesis. For the other parental strand, the lagging strand, DnaG synthesizes an RNA primer every few kilobases (kb). The https:// ensures that you are connecting to the Diabetes. Association of guanine nucleotide-linked succinate thiokinase with ketone body utilization", "Deficiency of the alpha subunit of succinate-coenzyme A ligase causes fatal infantile lactic acidosis with mitochondrial DNA depletion", Complex III/Coenzyme Q - cytochrome c reductase, Electron-transferring-flavoprotein dehydrogenase, Mitochondrial permeability transition pore. A thermodynamically constrained set of parameter values is identified for this candidate mechanism. CoA is removed from succinyl-CoA, producing free succinate; this is coupled with substrate-level . 2002;51:24962504. An official website of the United States government. Novel reaction of succinyl coenzyme A (Succinyl-CoA) synthetase Simulation results of mechanism A,, Figure 3. Adenylate kinase is a phosphotransferase enzyme that catalyzes the interconversion of the various adenosine phosphates. (B) ATP (and small amounts of UTP) synthesis by the Scs and subunits when [-32P]GTP is used as a phosphodonor in the presence of NDPII (ADP, CDP, and UDP). sharing sensitive information, make sure youre on a federal When [-32P]GTP was used as a phosphodonor and NDPII (ADP-CDP-UDP), ranging from 0 to 1,000 M, were used as substrates in the reaction mixture, predominantly ATP synthesis was observed, although small amounts of UTP were also detected (Fig. GTP is also important in polysaccharide synthesis since GDP-mannose, which is derived from mannose 1-phosphate and GTP (17), is often an intermediate in the synthesis of mannose (or its uronic acid mannuronate)-containing polysaccharides. This process often consists of metabolic pathways. It should be noted, however, that GTP-generating complexes of Ndk with proteins such as pyruvate kinase or Pra (4, 5, 24) involves the truncated 12-kDa form of Ndk, which is generated uniquely in the mucoid cells by the periplasmic retention of elastase (9, 12). [16] It has been demonstrated that the gene for SCS (sucCD) is transcribed along with the gene for -ketoglutarate dehydrogenase (sucAB) under the control of a promoter called sdhC, which is part of the succinate dehydrogenase operon. In particular, the interaction of the phosphoryl group with the power helix of each subunit suggests how the phosphohistidine or Pi would stabilize the aP-dimer. National Library of Medicine Accessibility describe two isoforms of succinyl-CoA synthetase in mammals, one that specifies synthesis of ATP, and one that synthesises GTP.[10]. In further refining this model for catalysis, we have shown that the monothiophosphorylated E. coli enzyme does not catalyse exchange of 18O from the beta, gamma-bridge to the beta-non-bridge position of ATP, indicating that the enzyme does not undergo even transient bis-phosphorylation. The two subunits only interact with a single unit, whereas the units interact with a single unit (to form the dimer) and the subunit of the other dimer. DnaG is a bacterial DNA primase and is encoded by the dnaG gene. Succinyl-CoA synthetase from P. aeruginosa utilizes GDP or ADP and synthesizes GTP or ATP, respectively, with radiolabeled Pi as a phosphodonor. The reaction was carried out in 20 l of TMD buffer with NDP concentrations ranging from 0 to 1 mM. [7] This histidine residue becomes phosphorylated during the succinate forming step in the reaction mechanism. The EcoRI-SalI fragment also contains the 3 region (129 bp) of the lpd gene. Succinyl coenzyme A synthetase (SCS, also known as succinyl-CoA synthetase or succinate thiokinase or succinate-CoA ligase) is an enzyme that catalyzes the reversible reaction of succinyl-CoA to succinate. Careers, Unable to load your collection due to an error. They serve as monomeric units of the nucleic acid polymers deoxyribonucleic acid (DNA) and ribonucleic acid (RNA), both of which are essential biomolecules within all life-forms on Earth. Rang J, Xia Z, Shuai L, Cao L, Liu Y, Li X, Xie J, Li Y, Hu S, Xie Q, Xia L. Microb Cell Fact. Simulation results of proposed BiBiUniUni, Figure A8. The sucCD operon (2,025 bp) is located 324 bp downstream from the lpd gene. Since the cells do not have a functional citric acid cycle, acidosis results because cells are forced to choose lactic acid production as the primary means of producing ATP. Antibacterial drugs that prevent phosphorylation of histidine, like the molecule LY26650, are potent inhibitors of bacterial SCSs. The 3 substrates of this enzyme are GTP, succinate, and CoA, whereas its 3 products are GDP, phosphate, and succinyl-CoA . Further, a 3.5-kb DNA fragment containing the sucCD hybridizing fragment was subcloned and sequenced. to a resolution of 2.10 A, with PDB code 1CQJ. Biochim. [3] However, Scs can freely interconvert ATP and succinate to form succinyl-CoA, ADP, and inorganic Pi for anabolic reactions as well, particularly during anaerobic growth (22). Succinyl Coenzyme A - an overview | ScienceDirect Topics [8] This leads researchers to believe that the enzyme must undergo a major change in conformation to bring the histidine to the grasp domain and facilitate the formation of the nucleoside triphosphate. Addition of dead-end binding(s) to, Figure 5. The gel was exposed to a phosphorimager cassette, and the amount of 32P bound to the Scs subunit was determined by using the PhosphorImager (STORM 860; Molecular Dynamics). An alternative mechanism proposed for pig heart GTP-specific SCS, Figure A7. Kelly C J, Cha S. Nucleotide specificity of succinate thiokinases from bacteria. An official website of the United States government. Before This arrangement is also. We demonstrate that, in the presence of GDP, Scs synthesizes GTP using ATP as the phosphodonor and, in the presence of ADP, Scs synthesizes ATP using GTP as a phosphodonor. Lu Q, Inouye M. Adenylate kinase complements nucleoside diphosphate kinase deficiency in nucleotide metabolism. It is unclear if such a complex plays any role in generating the GTP involved in alginate synthesis or if Scs is involved in residual alginate synthesis by the ndk knockout mutant cells. We have observed that the 16-kDa Ndk-Scs complex present in nonmucoid cells, synthesizes all three of the nucleoside triphosphates from a mixture of GDP, UDP, and CDP, whereas the 12-kDa Ndk-Scs complex specifically present in mucoid cell predominantly synthesizes GTP and UTP but not CTP. National Library of Medicine It is one of the building blocks needed for the synthesis of RNA during the transcription process. Succinyl-CoA - Wikipedia Chem. However, mammalian mitochondrial SCSs are active as dimers and do not form a heterotetramer. A 50 M concentration of NDPI (GDP-CDP-UDP) was used in all of the reactions in addition to [-32P]ATP as described previously (24). [11], SCS facilitates the flux of molecules into other metabolic pathways by controlling the interconversion between succinyl CoA and succinate. The Preparation, Properties, and Reactions of Succinyl Coenzyme A The substrate level phosphorylation catalyzed by Scs is known to involve adenine nucleotides in plants as well as in prokaryotes such as E. coli generating ATP during the operation of the TCA cycle (19, 20). Comprehensive discovery of novel structured noncoding RNAs in 26 bacterial genomes. (Fig.4B).4B). Blood cells, immune cells and intestinal bacteria as well as the intestinal mucosa also have a special need for nucleotides. In enzymology, a succinate-CoA ligase (ADP-forming) is an enzyme that catalyzes the chemical reaction, In enzymology, a succinateCoA ligase (GDP-forming) is an enzyme that catalyzes the chemical reaction. This is true when either ATP or succinyl-CoA and Pi are used to phosphorylate the enzyme. Addition of dead-end binding(s) to mechanisms A and E, Figure 6. variants are produced at different amounts in different tissues,[10] causing GTP or ATP substrate requirements. Labbe R. F., Kurumada T., Onisawa J. [18]. In Escherichia coli, Scs occurs predominantly in the A-form, where it exists as a heterotetramer consisting of 2 and 2 subunits. In this study, we show that in P. aeruginosa Scs allows the synthesis of both ATP and GTP from succinyl-CoA, Pi, and an equimolar mixture of ADP and GDP. As shown in the Fig. Joyce M. A. Ph. (Fig.4C).4C). The crystal structure of Succinyl-CoA synthetase alpha subunit (succinyl-CoA-binding isoform) was determined by Joyce et al. Autophosphorylation was carried out in the presence of various amounts of ADP or GDP and by using [-32P]ATP as the phosphodonor. Before Biosynthesis is usually synonymous with anabolism. Thus, synthesis of each molecule of alginate by mucoid P. aeruginosa requires the consumption of hundreds of molecules of GTP (4, 9). Simulation results of mechanism D,, Figure A5. We thank T. K. Misra for technical help on protein purification and Page Goodlove for DNA sequencing at the DNA Sequencing Facility of the University of Illinois at Urbana. Besides Ndk, other kinases, such as pyruvate kinase (24), adenylate kinase (15), or polyphosphate kinase (13), can substitute for Ndk, generating NTPs, including GTP. [10]. Seyfried TN, Arismendi-Morillo G, Mukherjee P, Chinopoulos C. iScience. Would you like email updates of new search results? [7] However, mammals possess different types of SCSs that are specific for either GTP (G-SCS) or ATP (A-SCS) and are native to different types of tissue within the organism. 2018 Jan-Dec;10:1759091418818261. doi: 10.1177/1759091418818261. [13] This is important because succinyl CoA is an intermediate necessary for porphyrin, heme,[14] and ketone body biosynthesis. This enzyme catalyzes the reversible formation of succinyl-Cob from succinate and Cob with the concomitant cleavage of ATP to ADP and Pi. A 3.5-kb EcoRI-SalI fragment was subcloned from pSU6 into pUC118 to generate pXL804. eCollection 2020 Nov 20. The loss of CTP synthesis by Scs, when complexed with the 12-kDa form of Ndk, is interesting. (Fig.7,7, lanes 9, 10, and 11). official website and that any information you provide is encrypted The lysed cells were pelleted at 15,000 rpm for 10 min at 4C, and the supernatant was filtered by using a 0.22-m (pore-size) filter. Genes Nutr. However, mammalian mitochondrial SCSs are active as dimers and do not form a heterotetramer. Clipboard, Search History, and several other advanced features are temporarily unavailable. (A) GTP synthesis by the Scs enzyme in the presence of 50 M ADP in each lane with various micromolar amounts of GDP in the reaction mixture as indicated. . Experimental data are same as in Figure 3. Succinyl coenzyme A synthetase from Escherichia coli does not exhibit a steady state kinetic pattern indicative of Mg+% E-P + succinate + CoA \ E + Pi + succinyl-CoA (2) "ping-pong" kinetics, despite the fact that catalytic participa- tion of a phosphorylated enzyme covalent intermediate has been established. Fatty acyl coenzyme A (CoA) synthetases are a class of enzymes involved in the activation of fatty acids through ligated high-energy CoA thioester bonds. It has been demonstrated that the membrane-associated 12-kDa nucleoside diphosphate (NDP) kinase (Ndk), in complex with pyruvate kinase or Pra (5, 24), specifically synthesizes GTP. 2016 Aug;72(Pt 8):912-21. doi: 10.1107/S2059798316010044. NCI CPTC Antibody Characterization Program. Succinyl Coenzyme A Synthetase from Escherichia coli - ResearchGate Unlike pyruvate kinase or Pra, which generates predominantly GTP in complexation with the 12-kDa form of Ndk (5, 24), Scs12-kDa-Ndk complexes generated both UTP and GTP.