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Escherichia coli K-12 substr. MG1655 Enzyme: glutamate-pyruvate aminotransferase



Gene: alaA Accession Numbers: G7184 (EcoCyc), b2290, ECK2284

Synonyms: yfbQ

Regulation Summary Diagram: ?

Subunit composition of glutamate-pyruvate aminotransferase = [AlaA]2
         glutamate-pyruvate aminotransferase = AlaA

Summary:
AlaA is one of three major alanine-synthesizing transaminases. AlaA and AlaC together account for 90% of glutamic-pyruvic transaminase (GPT) activity in the cell [Kim10].

A crystal structure of AlaA has been solved at 2.11 Å resolution. The structure shows a symmetric α2 homodimer typical of fold type I aminotransferases [PenaSoler14].

An alaA deletion strain has no growth defect, but an alaA avtA double mutant forms small colonies on agar plates. An alaA avtA alaC triple mutant has a slow growth phenotype in liquid medium. The defects of the double and triple mutants can be rescued by addition of alanine [Kim10, Yoneyama11]. Fitness and competitive growth experiments were performed under different growth conditions. Particularly under oxygen-limiting conditions, the doubling time of the ΔalaA strain in minimal media is increased compared to growth in rich media. Under competitive growth conditions, the ΔalaA mutation confers a disadvantage compared to wild type even in rich media [PenaSoler14]. alaA was identified in a screen for genes that reduce the lethal effects of stress. An alaA insertion mutant is more sensitive than wild type to mitomycin C and other stresses and less sensitive to 10% SDS [Han10a].

The alaA gene was first identified as a mutant with a leaky requirement for alanine or valine [Wang87].

alaA and yfbR may form an operon [Kim10].

Locations: cytosol

Map Position: [2,405,583 -> 2,406,800] (51.85 centisomes)
Length: 1218 bp / 405 aa

Molecular Weight of Polypeptide: 45.517 kD (from nucleotide sequence)

Molecular Weight of Multimer: 87.0 kD (experimental) [Kim10]

Unification Links: ASAP:ABE-0007560 , EchoBASE:EB3854 , EcoGene:EG14101 , EcoliWiki:b2290 , ModBase:P0A959 , OU-Microarray:b2290 , PortEco:yfbQ , Pride:P0A959 , Protein Model Portal:P0A959 , RefSeq:NP_416793 , RegulonDB:G7184 , SMR:P0A959 , String:511145.b2290 , Swiss-Model:P0A959 , UniProt:P0A959

Relationship Links: InterPro:IN-FAMILY:IPR004839 , InterPro:IN-FAMILY:IPR015421 , InterPro:IN-FAMILY:IPR015422 , InterPro:IN-FAMILY:IPR015424 , PDB:Structure:4CVQ , Pfam:IN-FAMILY:PF00155 , Prosite:IN-FAMILY:PS00105

In Paralogous Gene Group: 163 (5 members)

Gene-Reaction Schematic: ?

Genetic Regulation Schematic: ?

GO Terms:

Biological Process: GO:0006523 - alanine biosynthetic process Inferred from experiment [Kim10]
GO:0006974 - cellular response to DNA damage stimulus Inferred from experiment [Han10a]
GO:0019272 - L-alanine biosynthetic process from pyruvate Inferred from experiment [Yoneyama11, Kim10]
GO:0030632 - D-alanine biosynthetic process Inferred from experiment [Kim10]
GO:0046677 - response to antibiotic Inferred from experiment [Han10a]
GO:0009058 - biosynthetic process Inferred by computational analysis [GOA01a]
Molecular Function: GO:0004021 - L-alanine:2-oxoglutarate aminotransferase activity Inferred from experiment Inferred by computational analysis [GOA01, Kim10]
GO:0008483 - transaminase activity Inferred from experiment Inferred by computational analysis [UniProtGOA11a, Kim10]
GO:0030170 - pyridoxal phosphate binding Inferred from experiment Inferred by computational analysis [GOA01a, PenaSoler14]
GO:0042803 - protein homodimerization activity Inferred from experiment [Kim10]
GO:0003824 - catalytic activity Inferred by computational analysis [GOA01a]
GO:0016740 - transferase activity Inferred by computational analysis [UniProtGOA11a]
Cellular Component: GO:0005737 - cytoplasm Inferred by computational analysis [UniProtGOA11, UniProtGOA11a]
GO:0005829 - cytosol Inferred by computational analysis [DiazMejia09]

MultiFun Terms: metabolism biosynthesis of building blocks amino acids alanine

Essentiality data for alaA knockouts: ?

Growth Medium Growth? T (°C) O2 pH Osm/L Growth Observations
LB Lennox Yes 37 Aerobic 7   Yes [Baba06, Comment 1]
M9 medium with 1% glycerol Yes 37 Aerobic 7.2 0.35 Yes [Joyce06, Comment 2]
MOPS medium with 0.4% glucose Yes 37 Aerobic 7.2 0.22 Yes [Baba06, Comment 1]

Credits:
Created 31-Aug-2010 by Keseler I , SRI International
Last-Curated ? 15-Jul-2014 by Keseler I , SRI International


Enzymatic reaction of: glutamate-pyruvate aminotransferase

EC Number: 2.6.1.2

2-oxoglutarate + L-alanine <=> L-glutamate + pyruvate

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

This reaction is reversible.

In Pathways: superpathway of alanine biosynthesis , alanine biosynthesis II

Kinetic Parameters:

Substrate
Km (μM)
Citations
pyruvate
550.0
[Kim10]
L-alanine
4900.0
[Kim10]


Sequence Features

Feature Class Location Citations Comment
Amino-Acid-Sites-That-Bind 41
[UniProt10b]
UniProt: Substrate; via amide nitrogen; Non-Experimental Qualifier: by similarity;
Amino-Acid-Sites-That-Bind 179
[UniProt10b]
UniProt: Substrate; Non-Experimental Qualifier: by similarity;
N6-pyridoxal-phosphate-Lys-Modification 240
[UniProt11a]
UniProt: N6-(pyridoxal phosphate)lysine; Non-Experimental Qualifier: by similarity.
Amino-Acid-Sites-That-Bind 378
[UniProt10b]
UniProt: Substrate; Non-Experimental Qualifier: by similarity;


Gene Local Context (not to scale): ?

Transcription Units:

Notes:

History:
Markus Krummenacker on Tue Oct 14, 1997:
Gene object created from Blattner lab Genbank (v. M52) entry.


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

GOA01: GOA, MGI (2001). "Gene Ontology annotation based on Enzyme Commission mapping." Genomics 74;121-128.

GOA01a: GOA, DDB, FB, MGI, ZFIN (2001). "Gene Ontology annotation through association of InterPro records with GO terms."

Han10a: Han X, Dorsey-Oresto A, Malik M, Wang JY, Drlica K, Zhao X, Lu T (2010). "Escherichia coli genes that reduce the lethal effects of stress." BMC Microbiol 10;35. PMID: 20128927

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

Kim10: Kim SH, Schneider BL, Reitzer L (2010). "Genetics and regulation of the major enzymes of alanine synthesis in Escherichia coli." J Bacteriol 192(20);5304-11. PMID: 20729367

PenaSoler14: Pena-Soler E, Fernandez FJ, Lopez-Estepa M, Garces F, Richardson AJ, Quintana JF, Rudd KE, Coll M, Vega MC (2014). "Structural Analysis and Mutant Growth Properties Reveal Distinctive Enzymatic and Cellular Roles for the Three Major L-Alanine Transaminases of Escherichia coli." PLoS One 9(7);e102139. PMID: 25014014

UniProt10b: UniProt Consortium (2010). "UniProt version 2010-07 released on 2010-06-15 00:00:00." Database.

UniProt11a: UniProt Consortium (2011). "UniProt version 2011-11 released on 2011-11-22 00:00:00." Database.

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

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

Wang87: Wang MD, Buckley L, Berg CM (1987). "Cloning of genes that suppress an Escherichia coli K-12 alanine auxotroph when present in multicopy plasmids." J Bacteriol 169(12);5610-4. PMID: 2890623

Yoneyama11: Yoneyama H, Hori H, Lim SJ, Murata T, Ando T, Isogai E, Katsumata R (2011). "Isolation of a Mutant Auxotrophic for L-Alanine and Identification of Three Major Aminotransferases That Synthesize L-Alanine in Escherichia coli." Biosci Biotechnol Biochem 75(5);930-8. PMID: 21597182

Other References Related to Gene Regulation

Huerta03: Huerta AM, Collado-Vides J (2003). "Sigma70 promoters in Escherichia coli: specific transcription in dense regions of overlapping promoter-like signals." J Mol Biol 333(2);261-78. PMID: 14529615


Report Errors or Provide Feedback
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 Mon Dec 22, 2014, BIOCYC13A.