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MetaCyc Enzyme: GMP synthetase

Gene: guaA Accession Numbers: EG10420 (MetaCyc), b2507, ECK2503

Species: Escherichia coli K-12 substr. MG1655

Subunit composition of GMP synthetase = [GuaA]2
         GMP synthetase = GuaA

Summary:
GMP synthetase catalyzes the glutamine- or ammonia-dependent synthesis of GMP from XMP [Patel75].

GMP synthetase contains an N-terminal glutamine amide transfer (GAT) domain [Zalkin85a], a central ATP-pyrophosphatase (ATPP) domain, and a C-terminal dimerization domain [Tesmer96]. The GAT domain is thought to utilize glutamine to generate ammonia, which is then transferred to the ATPP domain via a substrate-protective channel or tunnel.

A protein consisting only of the ATPP and dimerization domains of GMP synthetase dimerizes in solution and has similar Km values for ATP, XMP, and ammonia as the full-length protein. However, the rate of catalysis using ammonia as a substrate is dramatically increased, possibly due to enhanced access to the active site [Abbott06].

A crystal stucture of GMP synthetase has been solved at 2.2 Å resolution [Tesmer96]. The enzyme from E. coli B is a dimer in solution [Sakamoto72], while preliminary analysis of the crystal structure of GMP synthetase showed a homotetrameric structure [Tesmer94].

Strains containing a guaA null mutation are auxotrophic for guanine [Lambden73, Joyce06].

A guaA-disrupted mutant was used to shut down the XMP to GMP reaction in a inosine-producing strain of E. coli [Shimaoka06].

Reviews: [Raushel99, Zalkin98]

Locations: cytosol

Map Position: [2,628,980 <- 2,630,557]

Molecular Weight of Polypeptide: 58.679 kD (from nucleotide sequence)

Molecular Weight of Multimer: 126 kD (experimental) [Sakamoto72]

pI: 5.46

Unification Links: ASAP:ABE-0008254 , CGSC:657 , DIP:DIP-9852N , EchoBASE:EB0415 , EcoGene:EG10420 , EcoliWiki:b2507 , ModBase:P04079 , OU-Microarray:b2507 , PortEco:guaA , Pride:P04079 , Protein Model Portal:P04079 , RefSeq:NP_417002 , RegulonDB:EG10420 , SMR:P04079 , String:511145.b2507 , UniProt:P04079

Relationship Links: InterPro:IN-FAMILY:IPR001674 , InterPro:IN-FAMILY:IPR004739 , InterPro:IN-FAMILY:IPR014729 , InterPro:IN-FAMILY:IPR017926 , InterPro:IN-FAMILY:IPR022310 , InterPro:IN-FAMILY:IPR022955 , InterPro:IN-FAMILY:IPR025777 , PDB:Structure:1GPM , Pfam:IN-FAMILY:PF00117 , Pfam:IN-FAMILY:PF00958 , Pfam:IN-FAMILY:PF02540 , Prosite:IN-FAMILY:PS51273 , Prosite:IN-FAMILY:PS51553

Gene-Reaction Schematic: ?

GO Terms:

Biological Process: GO:0006177 - GMP biosynthetic process Inferred from experiment Inferred by computational analysis [UniProtGOA12, UniProtGOA11, GOA06, GOA01, Lambden73]
GO:0006164 - purine nucleotide biosynthetic process Inferred by computational analysis [UniProtGOA11, GOA01]
GO:0006541 - glutamine metabolic process Inferred by computational analysis [UniProtGOA11, GOA06]
Molecular Function: GO:0003921 - GMP synthase activity Inferred from experiment [Abbott06]
GO:0003922 - GMP synthase (glutamine-hydrolyzing) activity Inferred from experiment Inferred by computational analysis [GOA06, GOA01a, GOA01, Deras99]
GO:0042802 - identical protein binding Inferred from experiment [Lasserre06]
GO:0042803 - protein homodimerization activity Inferred from experiment [Sakamoto72]
GO:0000166 - nucleotide binding Inferred by computational analysis [UniProtGOA11]
GO:0005524 - ATP binding Inferred by computational analysis [UniProtGOA11, GOA06, GOA01]
GO:0016462 - pyrophosphatase activity Inferred by computational analysis [GOA01]
GO:0016874 - ligase activity Inferred by computational analysis [UniProtGOA11]
Cellular Component: GO:0005829 - cytosol Inferred from experiment Inferred by computational analysis [DiazMejia09, Ishihama08, Lasserre06]

MultiFun Terms: metabolism biosynthesis of building blocks nucleotides purine biosynthesis
metabolism central intermediary metabolism nucleotide and nucleoside conversions

Credits:
Imported from EcoCyc 16-Sep-2014 by Paley S , SRI International


Enzymatic reaction of: xanthosine-5'-phosphate:L-glutamine amido-ligase (AMP-forming) (GMP synthetase)

Synonyms: GMP synthetase (glutamine-hydrolysing), glutamine amidotransferase, GMP synthetase, XMP aminase, GMPS

EC Number: 6.3.5.2

XMP + L-glutamine + ATP + H2O <=> L-glutamate + GMP + AMP + diphosphate + 2 H+

The reaction direction shown, that is, A + B ↔ C + D versus C + D ↔ A + B, is in accordance with the direction of enzyme catalysis.

The reaction is physiologically favored in the direction shown.

In Pathways: glutamine degradation I , superpathway of histidine, purine, and pyrimidine biosynthesis , superpathway of purine nucleotides de novo biosynthesis II , superpathway of guanosine nucleotides de novo biosynthesis II , superpathway of guanosine nucleotides de novo biosynthesis I , guanosine ribonucleotides de novo biosynthesis

Credits:
Imported from EcoCyc 16-Sep-2014 by Paley S , SRI International

Summary:
Early assays of the enzymatic activity (e.g. [Patel77], [Patel75], [Zalkin77], [Spector75]) were performed with enzyme purified from E. coli B 96, a purine-requiring derivative of E. coli B.

Cofactors or Prosthetic Groups: Mg2+ [vonderSaal85]

Inhibitors (Competitive): diphosphate [Spector75, Patel77] , N2-hydroxyguanosine 5'-monophosphate [Deras99]

Inhibitors (Irreversible): psicofuranine [Patel75, Patel77, vonderSaal85] , 6-diazo-5-oxonorleucine [Patel77, Chittur01]

Inhibitors (Unknown Mechanism): 3-bromopyruvate [Patel77] , 5,5'-dithio-bis-2-nitrobenzoate [Patel77] , tris [Patel77] , p-chloromercuribenzoate [Patel77] , acivicin [Chittur01]

Kinetic Parameters:

Substrate
Km (μM)
Citations
XMP
14.0
[Deras99]
L-glutamine
1000.0
[Chittur01]
ATP
1000.0
[Chittur01]

pH(opt): 8.3 [Patel75]


Enzymatic reaction of: xanthosine-5'-phosphate:ammonia ligase (AMP-forming) (GMP synthetase)

Synonyms: xanthosine-5'-phosphate ammonia ligase, GMP synthetase, XMP aminase, GMP synthetase (ammonia dependent), xanthosine 5'-phosphate aminase

XMP + ammonium + ATP <=> GMP + AMP + diphosphate + 2 H+

The reaction direction shown, that is, A + B ↔ C + D versus C + D ↔ A + B, is in accordance with the direction of enzyme catalysis.

The reaction is physiologically favored in the direction shown.

Credits:
Imported from EcoCyc 16-Sep-2014 by Paley S , SRI International

Summary:
Early assays of the enzymatic activity (e.g. [Patel77], [Patel75], [Zalkin77], [Spector75]) were performed with enzyme purified from E. coli B 96, a purine-requiring derivative of E. coli B. A variety of inhibitors of the enzyme were tested in [Spector75].

Inhibitors (Competitive): diphosphate [Spector75]

Inhibitors (Irreversible): 6-diazo-5-oxonorleucine [Patel77]

Inhibitors (Unknown Mechanism): decoyenine [Spector75] , adenosine [Spector75] , psicofuranine [Udaka63]

Kinetic Parameters:

Substrate
Km (μM)
Citations
XMP
58.0
[Spector74]


Sequence Features

Feature Class Location Citations Comment
Sequence-Conflict 3
[Tiedeman85a, UniProt10a]
Alternate sequence: N; UniProt: (in Ref. 1; AA sequence);
Conserved-Region 9 -> 207
[UniProt09]
UniProt: Glutamine amidotransferase type-1;
Active-Site 86
[UniProt10a]
UniProt: Nucleophile;
Active-Site 181
[UniProt10a]
Active-Site 183
[UniProt10a]
Conserved-Region 208 -> 400
[UniProt12]
UniProt: GMPS ATP-PPase.
Nucleotide-Phosphate-Binding-Region 235 -> 241
[UniProt10a]
UniProt: ATP;

History:
10/20/97 Gene b2507 from Blattner lab Genbank (v. M52) entry merged into EcoCyc gene EG10420; confirmed by SwissProt match.


References

Abbott06: Abbott JL, Newell JM, Lightcap CM, Olanich ME, Loughlin DT, Weller MA, Lam G, Pollack S, Patton WA (2006). "The Effects of Removing the GAT Domain from E. coli GMP Synthetase." Protein J 25;483-491. PMID: 17103135

Chittur01: Chittur SV, Klem TJ, Shafer CM, Davisson VJ (2001). "Mechanism for acivicin inactivation of triad glutamine amidotransferases." Biochemistry 40(4);876-87. PMID: 11170408

Deras99: Deras ML, Chittur SV, Davisson VJ (1999). "N2-hydroxyguanosine 5'-monophosphate is a time-dependent inhibitor of Escherichia coli guanosine monophosphate synthetase." Biochemistry 38(1);303-10. PMID: 9890911

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

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.

GOA06: GOA, SIB (2006). "Electronic Gene Ontology annotations created by transferring manual GO annotations between orthologous microbial proteins."

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

Lambden73: Lambden PR, Drabble WT (1973). "The gua operon of Escherichia coli K-12: evidence for polarity from guaB to guaA." J Bacteriol 115(3);992-1002. PMID: 4353875

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

Patel75: Patel N, Moyed HS, Kane JF (1975). "Xanthosine-5'-phosphate amidotransferase from Escherichia coli." J Biol Chem 250(7);2609-13. PMID: 235520

Patel77: Patel N, Moyed HS, Kane JF (1977). "Properties of xanthosine 5'-monophosphate-amidotransferase from Escherichia coli." Arch Biochem Biophys 1977;178(2);652-61. PMID: 189701

Raushel99: Raushel FM, Thoden JB, Holden HM (1999). "The amidotransferase family of enzymes: molecular machines for the production and delivery of ammonia." Biochemistry 38(25);7891-9. PMID: 10387030

Sakamoto72: Sakamoto N, Hatfield GW, Moyed HS (1972). "Physical properties and subunit structure of xanthosine 5'-phosphate aminase." J Biol Chem 247(18);5880-7. PMID: 4560421

Shimaoka06: Shimaoka M, Takenaka Y, Mihara Y, Kurahashi O, Kawasaki H, Matsui H (2006). "Effects of xapA and guaA disruption on inosine accumulation in Escherichia coli." Biosci Biotechnol Biochem 70(12);3069-72. PMID: 17151449

Spector74: Spector T, Miller RL, Fyfe JA, Krenitsky TA (1974). "GMP synthetase from Escherichia coli B-96. Interactions with substrate analogs." Biochim Biophys Acta 370(2);585-91. PMID: 4613384

Spector75: Spector T, Beacham LM (1975). "Guanosine monophosphate synthetase from Escherichia coli B-96. Inhibition by nucleosides." J Biol Chem 250(8);3101-7. PMID: 164459

Tesmer94: Tesmer JJ, Stemmler TL, Penner-Hahn JE, Davisson VJ, Smith JL (1994). "Preliminary X-ray analysis of Escherichia coli GMP synthetase: determination of anomalous scattering factors for a cysteinyl mercury derivative." Proteins 18(4);394-403. PMID: 8208731

Tesmer96: Tesmer JJ, Klem TJ, Deras ML, Davisson VJ, Smith JL (1996). "The crystal structure of GMP synthetase reveals a novel catalytic triad and is a structural paradigm for two enzyme families." Nat Struct Biol 3(1);74-86. PMID: 8548458

Tiedeman85a: Tiedeman AA, Smith JM, Zalkin H (1985). "Nucleotide sequence of the guaA gene encoding GMP synthetase of Escherichia coli K12." J Biol Chem 1985;260(15);8676-9. PMID: 3894345

Udaka63: Udaka S, Moyed HS (1963). "Inhibition of parental and mutant xanthosine 5'-phosphate aminases by psicofuranine." J Biol Chem 238;2797-803. PMID: 14063305

UniProt09: UniProt Consortium (2009). "UniProt version 15.8 released on 2009-10-01 00:00:00." Database.

UniProt10a: UniProt Consortium (2010). "UniProt version 2010-11 released on 2010-11-02 00:00:00." Database.

UniProt12: UniProt Consortium (2012). "UniProt version 2012-02 released on 2012-02-29 00:00:00." Database.

UniProtGOA11: 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."

vonderSaal85: von der Saal W, Crysler CS, Villafranca JJ (1985). "Positional isotope exchange and kinetic experiments with Escherichia coli guanosine-5'-monophosphate synthetase." Biochemistry 24(20);5343-50. PMID: 3907701

Zalkin77: Zalkin H, Truitt CD (1977). "Characterization of the glutamine site of Escherichia coli guanosine 5'-monophosphate synthetase." J Biol Chem 252(15);5431-6. PMID: 18463

Zalkin85a: Zalkin H, Argos P, Narayana SV, Tiedeman AA, Smith JM (1985). "Identification of a trpG-related glutamine amide transfer domain in Escherichia coli GMP synthetase." J Biol Chem 260(6);3350-4. PMID: 2982857

Zalkin98: Zalkin H, Smith JL (1998). "Enzymes utilizing glutamine as an amide donor." Adv Enzymol Relat Areas Mol Biol 72;87-144. PMID: 9559052


Report Errors or Provide Feedback
Please cite the following article in publications resulting from the use of MetaCyc: Caspi et al, Nucleic Acids Research 42:D459-D471 2014
Page generated by SRI International Pathway Tools version 18.5 on Sun Nov 23, 2014, biocyc14.