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Escherichia coli K-12 substr. MG1655 Enzyme: phosphoribosylaminoimidazole-succinocarboxamide synthase



Gene: purC Accession Numbers: EG10791 (EcoCyc), b2476, ECK2472

Synonyms: ade(g), ade

Regulation Summary Diagram: ?

Subunit composition of phosphoribosylaminoimidazole-succinocarboxamide synthase = [PurC]3

Summary:
PurC catalyzes the ligation of the carboxylate group of 5-amino-1-(5-phospho-D-ribosyl)imidazole-4-carboxylate (CAIR) to the amino group of aspartate [Zhang08f]. This is one of two reactions in purine metabolism that utilize aspartate and a ribonucleoside triphosphate, the other being adenylosuccinate synthetase (PurA). Their kinetic mechanisms are similar although they show only 16% amino acid sequence identity. For E. coli PurC a rapid equilibrium random ter-ter kinetic mechanism has been demonstrated [Nelson05].

The overproduction and purification of native [Meyer92] and hexahistidine-tagged recombinant [Nelson05] PurC have been reported.

A crystal structure of PurC in complex with ADP or ADP-CAIR has been determined [Ginder06].

Review: [Zhang08f]

Gene Citations: [Parker84, Tiedeman90, Meng90]

Locations: cytosol, membrane

Map Position: [2,594,927 <- 2,595,640] (55.93 centisomes)
Length: 714 bp / 237 aa

Molecular Weight of Polypeptide: 26.995 kD (from nucleotide sequence), 27.0 kD (experimental) [Meyer92 ]

Molecular Weight of Multimer: 76.0 kD (experimental) [Meyer92]

pI: 5.26

Unification Links: ASAP:ABE-0008156 , CGSC:343 , DIP:DIP-35900N , EchoBASE:EB0784 , EcoGene:EG10791 , EcoliWiki:b2476 , Mint:MINT-1257522 , ModBase:P0A7D7 , OU-Microarray:b2476 , PortEco:purC , PR:PRO_000023638 , Pride:P0A7D7 , Protein Model Portal:P0A7D7 , RefSeq:NP_416971 , RegulonDB:EG10791 , SMR:P0A7D7 , String:511145.b2476 , Swiss-Model:P0A7D7 , UniProt:P0A7D7

Relationship Links: InterPro:IN-FAMILY:IPR001636 , InterPro:IN-FAMILY:IPR013816 , InterPro:IN-FAMILY:IPR018236 , Panther:IN-FAMILY:PTHR11609 , PDB:Structure:2GQR , PDB:Structure:2GQS , Pfam:IN-FAMILY:PF01259 , Prosite:IN-FAMILY:PS01057 , Prosite:IN-FAMILY:PS01058

Gene-Reaction Schematic: ?

Genetic Regulation Schematic: ?

GO Terms:

Biological Process: GO:0006164 - purine nucleotide biosynthetic process Inferred by computational analysis [UniProtGOA11a, GOA01]
GO:0006189 - 'de novo' IMP biosynthetic process Inferred by computational analysis [UniProtGOA12]
Molecular Function: GO:0004639 - phosphoribosylaminoimidazolesuccinocarboxamide synthase activity Inferred from experiment Inferred by computational analysis [GOA01a, GOA01, Meyer92]
GO:0042802 - identical protein binding Inferred from experiment [Meyer92]
GO:0000166 - nucleotide binding Inferred by computational analysis [UniProtGOA11a]
GO:0005524 - ATP binding Inferred by computational analysis [UniProtGOA11a, GOA01]
GO:0016874 - ligase activity Inferred by computational analysis [UniProtGOA11a]
Cellular Component: GO:0005829 - cytosol Inferred from experiment Inferred by computational analysis [DiazMejia09, Ishihama08, Zhang07, LopezCampistrou05, Lasserre06]
GO:0016020 - membrane Inferred from experiment [Lasserre06]

MultiFun Terms: metabolism biosynthesis of building blocks nucleotides purine biosynthesis

Essentiality data for purC knockouts: ?

Growth Medium Growth? T (°C) O2 pH Osm/L Growth Observations
LB enriched Yes 37 Aerobic 6.95   Yes [Gerdes03, Comment 1]
LB Lennox Yes 37 Aerobic 7   Yes [Baba06, Comment 2]
M9 medium with 0.4% glucose No 37 Aerobic 7.2 0.27 No [Patrick07, Comment 3]
M9 medium with 1% glycerol No 37 Aerobic 7.2 0.35 No [Joyce06]
MOPS medium with 0.4% glucose No 37 Aerobic 7.2 0.22 No [Baba06, Comment 2]
No [Feist07, Comment 4]

Credits:
Last-Curated ? 16-Dec-2011 by Fulcher C , SRI International


Enzymatic reaction of: phosphoribosylaminoimidazole-succinocarboxamide synthase

Synonyms: SAICAR synthetase, 5'-phosphoribosyl-5-aminoimidazole-4-N-succinocarboxamide synthetase, phosphoribosylaminoimidazole succinocarboxamide synthetase

EC Number: 6.3.2.6

ATP + 5-amino-1-(5-phospho-D-ribosyl)imidazole-4-carboxylate + L-aspartate <=> ADP + 5'-phosphoribosyl-4-(N-succinocarboxamide)-5-aminoimidazole + phosphate + H+

The reaction direction shown, that is, A + B ↔ C + D versus C + D ↔ A + B, is in accordance with the Enzyme Commission system.

The reaction is physiologically favored in the direction shown.

Alternative Substrates for 5-amino-1-(5-phospho-D-ribosyl)imidazole-4-carboxylate: L-alanosine [Nelson05 ]

In Pathways: superpathway of histidine, purine, and pyrimidine biosynthesis , superpathway of purine nucleotides de novo biosynthesis II , inosine-5'-phosphate biosynthesis I

Summary:
For native PurC either a coupled assay utilizing pyruvate kinase/lactate dehydrogenase with monitoring the change in absorbance at 340 nm was used, or an assay utilizing radiolabeled aspartate was used. The Km value for ATP is for Mg-ATP [Meyer92].

For the recombinant, hexahistidine-tagged enzyme, IMP was a competitive inhibitor with respect to 5-amino-1-(5-phospho-D-ribosyl)imidazole-4-carboxylate (CAIR), and maleate was a competitive inhibitor with respect to aspartate [Nelson05].

Inhibitors (Competitive): IMP [Nelson05] , maleate [Nelson05]

Kinetic Parameters:

Substrate
Km (μM)
Citations
L-aspartate
1300.0
[Meyer92]
ATP
40.0
[Meyer92]
5-amino-1-(5-phospho-D-ribosyl)imidazole-4-carboxylate
36.0
[Meyer92]

pH(opt): 7.8 [Nelson05]


Gene Local Context (not to scale): ?

Transcription Unit:

Notes:

History:
10/20/97 Gene b2476 from Blattner lab Genbank (v. M52) entry merged into EcoCyc gene EG10791; confirmed by SwissProt match.


References

Baba06: Baba T, Ara T, Hasegawa M, Takai Y, Okumura Y, Baba M, Datsenko KA, Tomita M, Wanner BL, Mori H (2006). "Construction of Escherichia coli K-12 in-frame, single-gene knockout mutants: the Keio collection." Mol Syst Biol 2;2006.0008. PMID: 16738554

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

Feist07: Feist AM, Henry CS, Reed JL, Krummenacker M, Joyce AR, Karp PD, Broadbelt LJ, Hatzimanikatis V, Palsson BO (2007). "A genome-scale metabolic reconstruction for Escherichia coli K-12 MG1655 that accounts for 1260 ORFs and thermodynamic information." Mol Syst Biol 3;121. PMID: 17593909

Gerdes03: Gerdes SY, Scholle MD, Campbell JW, Balazsi G, Ravasz E, Daugherty MD, Somera AL, Kyrpides NC, Anderson I, Gelfand MS, Bhattacharya A, Kapatral V, D'Souza M, Baev MV, Grechkin Y, Mseeh F, Fonstein MY, Overbeek R, Barabasi AL, Oltvai ZN, Osterman AL (2003). "Experimental determination and system level analysis of essential genes in Escherichia coli MG1655." J Bacteriol 185(19);5673-84. PMID: 13129938

Ginder06: Ginder ND, Binkowski DJ, Fromm HJ, Honzatko RB (2006). "Nucleotide complexes of Escherichia coli phosphoribosylaminoimidazole succinocarboxamide synthetase." J Biol Chem 281(30);20680-8. PMID: 16687397

GOA01: GOA, DDB, FB, MGI, ZFIN (2001). "Gene Ontology annotation through association of InterPro records with GO terms."

GOA01a: GOA, MGI (2001). "Gene Ontology annotation based on Enzyme Commission mapping." Genomics 74;121-128.

Ishihama08: Ishihama Y, Schmidt T, Rappsilber J, Mann M, Hartl FU, Kerner MJ, Frishman D (2008). "Protein abundance profiling of the Escherichia coli cytosol." BMC Genomics 9;102. PMID: 18304323

Joyce06: Joyce AR, Reed JL, White A, Edwards R, Osterman A, Baba T, Mori H, Lesely SA, Palsson BO, Agarwalla S (2006). "Experimental and computational assessment of conditionally essential genes in Escherichia coli." J Bacteriol 188(23);8259-71. PMID: 17012394

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

LopezCampistrou05: Lopez-Campistrous A, Semchuk P, Burke L, Palmer-Stone T, Brokx SJ, Broderick G, Bottorff D, Bolch S, Weiner JH, Ellison MJ (2005). "Localization, annotation, and comparison of the Escherichia coli K-12 proteome under two states of growth." Mol Cell Proteomics 4(8);1205-9. PMID: 15911532

Meng90: Meng LM, Kilstrup M, Nygaard P (1990). "Autoregulation of PurR repressor synthesis and involvement of purR in the regulation of purB, purC, purL, purMN and guaBA expression in Escherichia coli." Eur J Biochem 1990;187(2);373-9. PMID: 2404765

Meyer92: Meyer E, Leonard NJ, Bhat B, Stubbe J, Smith JM (1992). "Purification and characterization of the purE, purK, and purC gene products: identification of a previously unrecognized energy requirement in the purine biosynthetic pathway." Biochemistry 1992;31(21);5022-32. PMID: 1534690

Nelson05: Nelson SW, Binkowski DJ, Honzatko RB, Fromm HJ (2005). "Mechanism of action of Escherichia coli phosphoribosylaminoimidazolesuccinocarboxamide synthetase." Biochemistry 44(2);766-74. PMID: 15641804

Parker84: Parker J (1984). "Identification of the purC gene product of Escherichia coli." J Bacteriol 157(3);712-7. PMID: 6365889

Patrick07: Patrick WM, Quandt EM, Swartzlander DB, Matsumura I (2007). "Multicopy suppression underpins metabolic evolvability." Mol Biol Evol 24(12);2716-22. PMID: 17884825

Tiedeman90: Tiedeman AA, DeMarini DJ, Parker J, Smith JM (1990). "DNA sequence of the purC gene encoding 5'-phosphoribosyl-5-aminoimidazole-4-N-succinocarboxamide synthetase and organization of the dapA-purC region of Escherichia coli K-12." J Bacteriol 172(10);6035-41. PMID: 2120198

UniProtGOA11a: UniProt-GOA (2011). "Gene Ontology annotation based on manual assignment of UniProtKB keywords in UniProtKB/Swiss-Prot entries."

UniProtGOA12: UniProt-GOA (2012). "Gene Ontology annotation based on UniPathway vocabulary mapping."

Zhang07: Zhang N, Chen R, Young N, Wishart D, Winter P, Weiner JH, Li L (2007). "Comparison of SDS- and methanol-assisted protein solubilization and digestion methods for Escherichia coli membrane proteome analysis by 2-D LC-MS/MS." Proteomics 7(4);484-93. PMID: 17309111

Zhang08f: Zhang Y, Morar M, Ealick SE (2008). "Structural biology of the purine biosynthetic pathway." Cell Mol Life Sci 65(23);3699-724. PMID: 18712276

Other References Related to Gene Regulation

Cho11: Cho BK, Federowicz SA, Embree M, Park YS, Kim D, Palsson BO (2011). "The PurR regulon in Escherichia coli K-12 MG1655." Nucleic Acids Res 39(15);6456-64. PMID: 21572102

He90a: He B, Shiau A, Choi KY, Zalkin H, Smith JM (1990). "Genes of the Escherichia coli pur regulon are negatively controlled by a repressor-operator interaction." J Bacteriol 1990;172(8);4555-62. PMID: 2198266


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Please cite the following article in publications resulting from the use of EcoCyc: Nucleic Acids Research 41:D605-12 2013
Page generated by SRI International Pathway Tools version 18.5 on Thu Dec 18, 2014, biocyc13.