|Gene:||sucC||Accession Numbers: EG10981 (MetaCyc), b0728, ECK0716|
Synonyms: β subunit
Species: Escherichia coli K-12 substr. MG1655
Component of: succinyl-CoA synthetase (extended summary available)
SucC is the β subunit of succinyl-CoA synthetase.
Trp45 and His50 are required for activity [Buck89], while Cys47 [Buck89] and Cys325 [Mann89] are not essential. Single mutations of tryptophan residues W43, W76 and W248 have little effect on enzyme activity. W76 and W248 may be part of the CoA binding site [Nishimura90]. Glu197 is essential for phosphorylation and dephosphorylation of the active site His246 in the α subunit [Fraser02].
SucC levels are elevated at low pH [Stancik, 2002].
|Map Position: [762,237 -> 763,403]|
Molecular Weight of Polypeptide: 41.393 kD (from nucleotide sequence), 41.0 kD (experimental) [Buck86 ]
Unification Links: ASAP:ABE-0002483 , CGSC:17845 , DIP:DIP-31852N , EchoBASE:EB0974 , EcoGene:EG10981 , EcoliWiki:b0728 , ModBase:P0A836 , OU-Microarray:b0728 , PortEco:sucC , PR:PRO_000024004 , Pride:P0A836 , Protein Model Portal:P0A836 , RefSeq:NP_415256 , RegulonDB:EG10981 , SMR:P0A836 , String:511145.b0728 , UniProt:P0A836
Relationship Links: InterPro:IN-FAMILY:IPR005809 , InterPro:IN-FAMILY:IPR005811 , InterPro:IN-FAMILY:IPR011761 , InterPro:IN-FAMILY:IPR013650 , InterPro:IN-FAMILY:IPR013815 , InterPro:IN-FAMILY:IPR013816 , InterPro:IN-FAMILY:IPR016102 , InterPro:IN-FAMILY:IPR017866 , Panther:IN-FAMILY:PTHR11815 , PDB:Structure:1CQI , PDB:Structure:1CQJ , PDB:Structure:1JKJ , PDB:Structure:1JLL , PDB:Structure:1SCU , PDB:Structure:2NU6 , PDB:Structure:2NU7 , PDB:Structure:2NU8 , PDB:Structure:2NU9 , PDB:Structure:2NUA , PDB:Structure:2SCU , Pfam:IN-FAMILY:PF00549 , Pfam:IN-FAMILY:PF08442 , Prosite:IN-FAMILY:PS01217 , Prosite:IN-FAMILY:PS50975
|Biological Process:||GO:0006099 - tricarboxylic acid cycle
[UniProtGOA12, UniProtGOA11, GOA06, Buck86]
GO:0008152 - metabolic process [GOA01]
|Molecular Function:||GO:0004775 - succinate-CoA ligase (ADP-forming) activity
[GOA06, GOA01a, Buck86]
GO:0005515 - protein binding [Lasserre06, Arifuzzaman06]
GO:0000166 - nucleotide binding [UniProtGOA11]
GO:0000287 - magnesium ion binding [GOA06]
GO:0003824 - catalytic activity [GOA01]
GO:0005524 - ATP binding [UniProtGOA11, GOA06, GOA01]
GO:0016874 - ligase activity [UniProtGOA11]
GO:0030145 - manganese ion binding [GOA06]
GO:0046872 - metal ion binding [UniProtGOA11, GOA01]
|Cellular Component:||GO:0005737 - cytoplasm
GO:0005829 - cytosol [DiazMejia09, Ishihama08]
GO:0009361 - succinate-CoA ligase complex (ADP-forming) [Wolodko94]
|MultiFun Terms:||metabolism → energy metabolism, carbon → TCA cycle|
Subunit of: succinyl-CoA synthetase
Species: Escherichia coli K-12 substr. MG1655
Succinyl-CoA synthetase catalyzes the only reaction of the TCA cycle that employs substrate-level phosphorylation. The reaction is reversible and is thought to proceed in three partial reactions via enzyme-bound succinyl-phosphate and a phosphorylated enzyme intermediate, where a histidine residue of the α subunit is transiently phosphorylated. Many biochemical studies were done with the enzyme purified from E. coli Crooks strain (see citations in the Enzymatic Reaction summary), while mutant enzymes were obtained from the genes of a K-12-derived strain.
A hybrid enzyme containing one wild-type and one catalytically inactive α subunit retains significant enzymatic activity, arguing for the presence of two independently active heterodimers [Mann91]. Crystal structures of the enzyme from various E. coli sources have been solved [Wolodko84, Wolodko94, Fraser99b, Joyce00, Fraser02, Hidber07]. The α2β2 heterotetramer is composed of two αβ heterodimers [Wolodko94]. Two possible alternative interpretations of the dimer-dimer interface were resolved by generating site-directed mutants that exist and are catalytically active as αβ heterodimers [Bailey99].
Overlap of the sucC and sucD open reading frames suggests translational coupling [Buck85].
A sucCD insertion mutant does not grow aearobically on acetate or α-ketoglutarate or anaerobically on glucose or succinate as carbon sources. Expression of sucCD is higher under aerobic than anaerobic conditions and is induced by growth on acetate [MatJan89a]. A sucCD deletion mutant shows higher acetate accumulation and lower biomass formation than wild type on glucose minimal medium and can not re-utilize acetate that is secreted during growth on glucose [Veit07]. sucCD and sucAB are mutually essential. Both sets of genes encode an enzyme that can produce succinyl-CoA, an essential precursor for peptidoglycan biosynthesis; deletion of either set alone results in a longer lag phase, while cells where both sets are deleted are not viable [Yu06b].
|Molecular Function:||GO:0004775 - succinate-CoA ligase (ADP-forming) activity [Luo91]|
Enzymatic reaction of: succinyl-CoA synthetase
Synonyms: succinate thiokinase
EC Number: 126.96.36.199
The reaction direction shown, that is, A + B ↔ C + D versus C + D ↔ A + B, is in accordance with the direction in which it was curated.
This reaction is reversible.
Initial studies of the enzyme were done in unidentified strains [Kreil64, Nishimura65, Nishimura67] and E. coli (Crookes strain), e.g. [Ramaley67, Sedmak68, Pearson75, Bridger68, Moffet72, Pearson75a, Grinnell69a, Lam76, Buttlaire77, Collier78, Collier79, Bild80, Vogel82, Prasad82, Prasad83, Nishimura83a, Prasad83a, Wolodko83, Nishimura84, Nishimura84a, Wolodko80a, Williams87a, Khan88a, Kelly77].
The enzyme can utilize some alternative substrates with low efficiency [Nolte13].
Cofactors or Prosthetic Groups: Mg2+
|Conserved-Region||9 -> 244|
|Nucleotide-Phosphate-Binding-Region||35 -> 108|
10/20/97 Gene b0728 from Blattner lab Genbank (v. M52) entry merged into EcoCyc gene EG10981; confirmed by SwissProt match.
Arifuzzaman06: Arifuzzaman M, Maeda M, Itoh A, Nishikata K, Takita C, Saito R, Ara T, Nakahigashi K, Huang HC, Hirai A, Tsuzuki K, Nakamura S, Altaf-Ul-Amin M, Oshima T, Baba T, Yamamoto N, Kawamura T, Ioka-Nakamichi T, Kitagawa M, Tomita M, Kanaya S, Wada C, Mori H (2006). "Large-scale identification of protein-protein interaction of Escherichia coli K-12." Genome Res 16(5);686-91. PMID: 16606699
Bailey99: Bailey DL, Fraser ME, Bridger WA, James MN, Wolodko WT (1999). "A dimeric form of Escherichia coli succinyl-CoA synthetase produced by site-directed mutagenesis." J Mol Biol 285(4);1655-66. PMID: 9917403
Bild80: Bild GS, Janson CA, Boyer PD (1980). "Subunit interaction during catalysis. ATP modulation of catalytic steps in the succinyl-CoA synthetase reaction." J Biol Chem 255(17);8109-15. PMID: 6997289
Bridger87: Bridger WA, Wolodko WT, Henning W, Upton C, Majumdar R, Williams SP (1987). "The subunits of succinyl-coenzyme A synthetase--function and assembly." Biochem Soc Symp 1987;54;103-11. PMID: 3332988
Buck89: Buck D, Guest JR (1989). "Overexpression and site-directed mutagenesis of the succinyl-CoA synthetase of Escherichia coli and nucleotide sequence of a gene (g30) that is adjacent to the suc operon." Biochem J 1989;260(3);737-47. PMID: 2548486
Buttlaire77: Buttlaire DH, Chon M (1977). "Interactions of phospho- and dephosphosuccinyl coenzyme A synthetase with manganous ion and substrates. Studies of manganese complexes by NMR relaxation rates of water protons." J Biol Chem 252(6);1957-64. PMID: 321448
Collier78: Collier GE, Nishimura JS (1978). "Affinity labeling of succinyl-CoA synthetase from porcine heart and Escherichia coli with oxidized coenzyme A disulfide." J Biol Chem 253(14);4938-43. PMID: 353044
DiazMejia09: Diaz-Mejia JJ, Babu M, Emili A (2009). "Computational and experimental approaches to chart the Escherichia coli cell-envelope-associated proteome and interactome." FEMS Microbiol Rev 33(1);66-97. PMID: 19054114
Fong92: Fong G, Bridger WA (1992). "Folding and assembly of the Escherichia coli succinyl-CoA synthetase heterotetramer without participation of molecular chaperones." Biochemistry 31(24);5661-4. PMID: 1351743
Fraser02: Fraser ME, Joyce MA, Ryan DG, Wolodko WT (2002). "Two glutamate residues, Glu 208 alpha and Glu 197 beta, are crucial for phosphorylation and dephosphorylation of the active-site histidine residue in succinyl-CoA synthetase." Biochemistry 41(2);537-46. PMID: 11781092
Grinnell69a: Grinnell FL, Nishimura JS (1969). "Succinate thiokinase of Escherichia coli. Purification, phosphorylation of the enzyme, and exchange reactions catalyzed by the enzyme." Biochemistry 8(2);562-8. PMID: 4240087
Hidber07: Hidber E, Brownie ER, Hayakawa K, Fraser ME (2007). "Participation of Cys123alpha of Escherichia coli succinyl-CoA synthetase in catalysis." Acta Crystallogr D Biol Crystallogr 63(Pt 8);876-84. PMID: 17642514
Joyce00: Joyce MA, Fraser ME, James MN, Bridger WA, Wolodko WT (2000). "ADP-binding site of Escherichia coli succinyl-CoA synthetase revealed by x-ray crystallography." Biochemistry 39(1);17-25. PMID: 10625475
Joyce99: Joyce MA, Fraser ME, Brownie ER, James MN, Bridger WA, Wolodko WT (1999). "Probing the nucleotide-binding site of Escherichia coli succinyl-CoA synthetase." Biochemistry 38(22);7273-83. PMID: 10353839
Lam76: Lam YF, Bridger WA, Kotowycz G (1976). "Nuclear magnetic resonance relaxation time studies on the manganese(II) ion complex with succinyl coenzyme A synthetase from Escherichia coli." Biochemistry 15(21);4742-8. PMID: 788782
Lasserre06: Lasserre JP, Beyne E, Pyndiah S, Lapaillerie D, Claverol S, Bonneu M (2006). "A complexomic study of Escherichia coli using two-dimensional blue native/SDS polyacrylamide gel electrophoresis." Electrophoresis 27(16);3306-21. PMID: 16858726
Luo91: Luo GX, Nishimura JS (1991). "Site-directed mutagenesis of Escherichia coli succinyl-CoA synthetase. Histidine 142 alpha is a facilitative catalytic residue." J Biol Chem 266(31);20781-5. PMID: 1939128
Luo92: Luo GX, Nishimura JS (1992). "Adenosine 5'-tetraphosphate is synthesized by the histidine alpha 142----asparagine mutant of Escherichia coli succinyl-CoA synthetase." J Biol Chem 267(14);9516-20. PMID: 1577794
Mann89: Mann CJ, Hardies SC, Nishimura JS (1989). "Site-directed mutagenesis of Escherichia coli succinyl-CoA synthetase. beta-Cys325 is a nonessential active site residue." J Biol Chem 264(3);1457-60. PMID: 2643599
Mann91: Mann CJ, Mitchell T, Nishimura JS (1991). "Phosphorylation and formation of hybrid enzyme species test the "half of sites" reactivity of Escherichia coli succinyl-CoA synthetase." Biochemistry 30(6);1497-503. PMID: 1993168
MatJan89a: Mat-Jan F, Williams CR, Clark DP (1989). "Anaerobic growth defects resulting from gene fusions affecting succinyl-CoA synthetase in Escherichia coli K12." Mol Gen Genet 215(2);276-80. PMID: 2496297
Moffet72: Moffet FJ, Wang T, Bridger WA (1972). "Succinyl coenzyme A synthetase of Escherichia coli. Effects of phosphoenzyme formation and of substrate binding on the reactivity and stability of the enzyme." J Biol Chem 247(24);8139-44. PMID: 4565676
Nishimura65: Nishimura JS, Meister A (1965). "Evidence for Succinyl Phosphate as an Enzyme-bound Intermediate in the Reaction Catalyzed by Succinyl Coenzyme A Synthetase." Biochemistry 4(7);1457-1462.
Nishimura83a: Nishimura JS, Mitchell T, Collier GE, Matula JM, Ball DJ (1983). "Affinity labeling of succinyl-CoA synthetase from Escherichia coli by the 2',3'-dialdehyde derivative of adenosine 5'-diphosphate." Eur J Biochem 136(1);83-7. PMID: 6352264
Nishimura84: Nishimura JS, Mitchell T (1984). "Escherichia coli succinyl coenzyme A synthetase. Inhibition of ATP-stimulated succinate----succinyl coenzyme A exchange at low succinyl coenzyme A concentrations by an ADP trap." J Biol Chem 259(4);2144-8. PMID: 6365903
Nishimura84a: Nishimura JS, Mitchell T (1984). "Adenosine 5'-O-(3-thio)triphosphate, a substrate and potent inhibitor of Escherichia coli succinyl-CoA synthetase. Additional evidence for a cooperative alternating-sites mechanism." J Biol Chem 259(15);9642-5. PMID: 6378911
Nishimura90: Nishimura JS, Mann CJ, Ybarra J, Mitchell T, Horowitz PM (1990). "Intrinsic fluorescence of succinyl-CoA synthetase and four tryptophan mutants. Tryptophan 76 and tryptophan 248 of the beta-subunit are responsive to CoA binding." Biochemistry 29(4);862-5. PMID: 2340278
Nishimura93: Nishimura JS, Ybarra J, Mann CJ, Mitchell T (1993). "Sensitivity of Escherichia coli succinyl-CoA mutants at Trp beta 76 to clostripain and to trypsin. ADP and ATP protect against cleavage by clostripain at Arg beta 80." J Biol Chem 268(18);13717-22. PMID: 8514803
Nolte13: Nolte JC, Schurmann M, Schepers CL, Vogel E, Wubbeler JH, Steinbuchel A (2014). "Novel characteristics of succinate coenzyme A (Succinate-CoA) ligases: conversion of malate to malyl-CoA and CoA-thioester formation of succinate analogues in vitro." Appl Environ Microbiol 80(1);166-76. PMID: 24141127
OConnorMcCourt85: O'Connor-McCourt MD, Bridger WA (1985). "Chemical modification and hybrid enzyme formation as probes of the active site and subunit interactions in Escherichia coli succinyl-CoA synthetase." Can J Biochem Cell Biol 63(1);57-63. PMID: 3886104
Pearson75: Pearson PH, Bridger WA (1975). "Isolation of the alpha and beta subunits of Escherichia coli succinyl coenzyme A synthetase and their recombination into active enzyme." J Biol Chem 250(12);4451-5. PMID: 1095571
Pearson75a: Pearson PH, Bridger WA (1975). "Catalysis of a step of the overall reaction by the alpha subunit of Escherichia coli succinyl coenzyme A synthetase." J Biol Chem 250(21);8524-9. PMID: 1104606
Prasad82: Prasad AR, Nishimura JS, Horowitz PM (1982). "Fluorescence detection of increased local flexibility induced by coenzyme A in succinyl-coA synthetase from Escherichia coli." Biochemistry 21(21);5142-7. PMID: 6756468
Prasad83: Prasad AR, Nishimura JS, Horowitz PM (1983). "A study of the quenching of the intrinsic fluorescence of succinyl-CoA synthetase from Escherichia coli by acrylamide, iodide, and coenzyme A." Biochemistry 22(18);4272-5. PMID: 6354251
Prasad83a: Prasad AR, Ybarra J, Nishimura JS (1983). "Chemical modification of Escherichia coli succinyl-CoA synthetase with the adenine nucleotide analogue 5'-p-fluorosulphonylbenzoyladenosine." Biochem J 215(3);513-8. PMID: 6362660
Ramaley67: Ramaley RF, Bridger WA, Moyer RW, Boyer PD (1967). "The preparation, properties, and reactions of succinyl coenzyme A synthetase and its phosphorylated form." J Biol Chem 242(19);4287-98. PMID: 4863737
Schauder87a: Schauder B, Blocker H, Frank R, McCarthy JE (1987). "Inducible expression vectors incorporating the Escherichia coli atpE translational initiation region." Gene 52(2-3);279-83. PMID: 3038690
Sedmak68: Sedmak J, Ramaley R (1968). "Identification of succinyl-CoA synthetase as the major phosphorylated protein formed from ATP in Escherichia coli extracts." Biochim Biophys Acta 170(2);440-2. PMID: 4885687
Stancik, 2002: Stancik LM, Stancik DM, Schmidt B, Barnhart DM, Yoncheva YN, Slonczewski JL (2002). "pH-dependent expression of periplasmic proteins and amino acid catabolism in Escherichia coli." J Bacteriol 184(15);4246-58. PMID: 12107143
Thomas74: Thomas EL (1974). "Studies on the metabolism of ATP by isolated bacterial membranes: role of succinyl CoA synthetase in diglyceride kinase activity." Arch Biochem Biophys 163(2);530-6. PMID: 4370406
Veit07: Veit A, Polen T, Wendisch VF (2007). "Global gene expression analysis of glucose overflow metabolism in Escherichia coli and reduction of aerobic acetate formation." Appl Microbiol Biotechnol 74(2);406-21. PMID: 17273855
Vogel82: Vogel HJ, Bridger WA, Sykes BD (1982). "Frequency-dependent phosphorus-31 nuclear magnetic resonance studies of the phosphohistidine residue to succinyl-CoA synthetase and the phosphoserine residue of glycogen phosphorylase a." Biochemistry 21(6);1126-32. PMID: 6803832
Williams87a: Williams SP, Bridger WA (1987). "Positional oxygen isotope exchange as a probe for the mechanism of catalysis by Escherichia coli succinyl coenzyme A synthetase." Biochemistry 26(14);4483-7. PMID: 3311148
Wolodko80a: Wolodko WT, Brownie ER, Bridger WA (1980). "Subunits of succinyl-coenzyme A synthetase: coordination of production in Escherichia coli and discovery of a factor that precludes refolding." J Bacteriol. 143(1): 231-237. PMID: 6995430
Wolodko83: Wolodko WT, Brownie ER, O'Connor MD, Bridger WA (1983). "Thiophosphorylation as a probe for subunit interactions in Escherichia coli succinyl coenzyme A synthetase. Further evidence for catalytic cooperativity and substrate synergism." J Biol Chem 258(23);14116-9. PMID: 6358215
Wolodko86: Wolodko WT, Kay CM, Bridger WA (1986). "Active enzyme sedimentation, sedimentation velocity, and sedimentation equilibrium studies of succinyl-CoA synthetases of porcine heart and Escherichia coli." Biochemistry 25(19);5420-5. PMID: 3535876
Wolodko87: Wolodko WT, Bridger WA (1987). "Studies of the process of renaturation and assembly of Escherichia coli succinyl-CoA synthetase from its alpha and beta subunits." Biochem Cell Biol 65(5);452-7. PMID: 3304349
Wolodko94: Wolodko WT, Fraser ME, James MN, Bridger WA (1994). "The crystal structure of succinyl-CoA synthetase from Escherichia coli at 2.5-A resolution." J Biol Chem 269(14);10883-90. PMID: 8144675
Ybarra86: Ybarra J, Prasad AR, Nishimura JS (1986). "Chemical modification of tryptophan residues in Escherichia coli succinyl-CoA synthetase. Effect on structure and enzyme activity." Biochemistry 25(22);7174-8. PMID: 3542020
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