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Escherichia coli K-12 substr. MG1655 Enzyme: asparagine synthetase A



Gene: asnA Accession Numbers: EG10091 (EcoCyc), b3744, ECK3738

Regulation Summary Diagram: ?

Subunit composition of asparagine synthetase A = [AsnA]2

Summary:
Asparagine synthetase A (AsnA) is one of two asparagine synthetases in E. coli, catalyzing the ammonia-dependent conversion of aspartate to asparagine.

AsnA is the more active of two asparagine synthetase activities in E. coli [Humbert80, Felton80].

Asparagine synthetase A comprises a dimer of AsnA monomers [Cedar69, Sugiyama92, Nakamura81]. A crystal structure of AsnA has been determined to 2.5 Å resolution [Nakatsu98].

Locations: cytosol

Map Position: [3,925,178 -> 3,926,170] (84.6 centisomes)
Length: 993 bp / 330 aa

Molecular Weight of Polypeptide: 36.651 kD (from nucleotide sequence)

Molecular Weight of Multimer: 80 kD (experimental) [Cedar69]

pI: 5.6 [Burland93], 5.5 [Cedar69]

Unification Links: ASAP:ABE-0012242 , CGSC:995 , DIP:DIP-9176N , EchoBASE:EB0089 , EcoGene:EG10091 , EcoliWiki:b3744 , ModBase:P00963 , OU-Microarray:b3744 , PortEco:asnA , PR:PRO_000022167 , Pride:P00963 , Protein Model Portal:P00963 , RefSeq:NP_418200 , RegulonDB:EG10091 , SMR:P00963 , String:511145.b3744 , UniProt:P00963

Relationship Links: InterPro:IN-FAMILY:IPR004618 , InterPro:IN-FAMILY:IPR006195 , PDB:Structure:11AS , PDB:Structure:12AS , Pfam:IN-FAMILY:PF03590 , Prosite:IN-FAMILY:PS50862

Gene-Reaction Schematic: ?

Genetic Regulation Schematic: ?

GO Terms:

Biological Process: GO:0006529 - asparagine biosynthetic process Inferred from experiment Inferred by computational analysis [UniProtGOA11, GOA01, RAVEL62]
GO:0006974 - cellular response to DNA damage stimulus Inferred from experiment [Khil02]
GO:0006418 - tRNA aminoacylation for protein translation Inferred by computational analysis [GOA01]
GO:0008652 - cellular amino acid biosynthetic process Inferred by computational analysis [UniProtGOA11]
GO:0070981 - L-asparagine biosynthetic process Inferred by computational analysis [UniProtGOA12]
Molecular Function: GO:0004071 - aspartate-ammonia ligase activity Inferred from experiment Inferred by computational analysis [GOA01a, GOA01, Cedar69]
GO:0000166 - nucleotide binding Inferred by computational analysis [UniProtGOA11]
GO:0004812 - aminoacyl-tRNA ligase activity Inferred by computational analysis [GOA01]
GO:0005524 - ATP binding Inferred by computational analysis [UniProtGOA11]
GO:0016874 - ligase activity Inferred by computational analysis [UniProtGOA11]
Cellular Component: GO:0005829 - cytosol Inferred from experiment Inferred by computational analysis [DiazMejia09, Ishihama08, LopezCampistrou05]
GO:0005737 - cytoplasm Inferred by computational analysis [UniProtGOA11a, UniProtGOA11, GOA01]

MultiFun Terms: metabolism biosynthesis of building blocks amino acids asparagine
metabolism carbon utilization amino acids

Essentiality data for asnA 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 1% glycerol Yes 37 Aerobic 7.2 0.35 Yes [Joyce06, Comment 3]
MOPS medium with 0.4% glucose Yes 37 Aerobic 7.2 0.22 Yes [Baba06, Comment 2]
Yes [Feist07, Comment 4]

Credits:
Last-Curated ? 10-Jan-2007 by Shearer A , SRI International
Revised 23-May-2011 by Brito D


Enzymatic reaction of: asparagine synthetase

Synonyms: aspartate-ammonia ligase

EC Number: 6.3.1.1

L-aspartate + ammonium + ATP <=> L-asparagine + AMP + diphosphate + 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 irreversible in the direction shown.

In Pathways: superpathway of asparagine biosynthesis , superpathway of aspartate and asparagine biosynthesis; interconversion of aspartate and asparagine , asparagine biosynthesis II

Summary:
This reaction is subject to competitive inhibition by ammonia (Ki 0.5mM), asparagine (Ki 0.12mM), and AMP (Ki 1.5mM) [Cedar69a].

Cofactors or Prosthetic Groups: Mg2+ [Cedar69]

Inhibitors (Competitive): AMP [Cedar69a, Comment 5] , L-asparagine [Cedar69a, Comment 6] , ammonia [Cedar69a]

Primary Physiological Regulators of Enzyme Activity: AMP , L-asparagine , ammonia

Kinetic Parameters:

Substrate
Km (μM)
Citations
ATP
270.0
[Nakatsu96, BRENDA14]

pH(opt): 8.4 [Cedar69]


Gene Local Context (not to scale): ?

Transcription Units:

Notes:

History:
10/20/97 Gene b3744 from Blattner lab Genbank (v. M52) entry merged into EcoCyc gene EG10091; 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

BRENDA14: BRENDA team (2014). "Imported from BRENDA version existing on Aug 2014." http://www.brenda-enzymes.org.

Burland93: Burland V, Plunkett G, Daniels DL, Blattner FR (1993). "DNA sequence and analysis of 136 kilobases of the Escherichia coli genome: organizational symmetry around the origin of replication." Genomics 1993;16(3);551-61. PMID: 7686882

Cedar69: Cedar H, Schwartz JH (1969). "The asparagine synthetase of Escherhic coli. I. Biosynthetic role of the enzyme, purification, and characterization of the reaction products." J Biol Chem 1969;244(15);4112-21. PMID: 4895361

Cedar69a: Cedar H, Schwartz JH (1969). "The asparagine synthetase of Escherichia coli. II. Studies on mechanism." J Biol Chem 1969;244(15);4122-7. PMID: 4895362

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

Felton80: Felton J, Michaelis S, Wright A (1980). "Mutations in two unlinked genes are required to produce asparagine auxotrophy in Escherichia coli." J Bacteriol 142(1);221-8. PMID: 6102983

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

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.

Humbert80: Humbert R, Simoni RD (1980). "Genetic and biomedical studies demonstrating a second gene coding for asparagine synthetase in Escherichia coli." J Bacteriol 1980;142(1);212-20. PMID: 6102982

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

Khil02: Khil PP, Camerini-Otero RD (2002). "Over 1000 genes are involved in the DNA damage response of Escherichia coli." Mol Microbiol 44(1);89-105. PMID: 11967071

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

Nakamura81: Nakamura M, Yamada M, Hirota Y, Sugimoto K, Oka A, Takanami M (1981). "Nucleotide sequence of the asnA gene coding for asparagine synthetase of E. coli K-12." Nucleic Acids Res 1981;9(18);4669-76. PMID: 6117826

Nakatsu96: Nakatsu T, Kato H, Oda J (1996). "Crystallization and preliminary crystallographic study of asparagine synthetase from Escherichia coli." Acta Crystallogr D Biol Crystallogr 52(Pt 3);604-6. PMID: 15299693

Nakatsu98: Nakatsu T, Kato H, Oda J (1998). "Crystal structure of asparagine synthetase reveals a close evolutionary relationship to class II aminoacyl-tRNA synthetase." Nat Struct Biol 5(1);15-9. PMID: 9437423

RAVEL62: RAVEL JM, NORTON SJ, HUMPHREYS JS, SHIVE W (1962). "Asparagine biosynthesis in Lactobacillus arabinosus and its control by asparagine through enzyme inhibition and repression." J Biol Chem 237;2845-9. PMID: 14490631

Sugiyama92: Sugiyama A, Kato H, Nishioka T, Oda J (1992). "Overexpression and purification of asparagine synthetase from Escherichia coli." Biosci Biotechnol Biochem 56(3);376-9. PMID: 1369484

UniProtGOA11: UniProt-GOA (2011). "Gene Ontology annotation based on manual assignment of UniProtKB keywords in UniProtKB/Swiss-Prot entries."

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

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

Other References Related to Gene Regulation

Kolling85: Kolling R, Lother H (1985). "AsnC: an autogenously regulated activator of asparagine synthetase A transcription in Escherichia coli." J Bacteriol 1985;164(1);310-5. PMID: 2864330

Poggio02: Poggio S, Domeinzain C, Osorio A, Camarena L (2002). "The nitrogen assimilation control (Nac) protein represses asnC and asnA transcription in Escherichia coli." FEMS Microbiol Lett 206(2);151-6. PMID: 11814655

Suzuki03: Suzuki M (2003). "The DNA-binding specificity of eubacterial and archaeal FFRPs. ." Proceedings of the Japan Academy, Series B79B(7); 213-222.

Tobia85: Tobia AJ, Couri D, Sagone A (1985). "The effects of the quinone type drugs on hydroxyl radical (OH.) production by rat liver microsomes." J Toxicol Environ Health 15(2);265-77. PMID: 3925152


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Please cite the following article in publications resulting from the use of EcoCyc: Nucleic Acids Research 41:D605-12 2013
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