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Escherichia coli K-12 substr. MG1655 Enzyme: homoserine O-succinyltransferase



Gene: metA Accession Numbers: EG10581 (EcoCyc), b4013, ECK4005

Regulation Summary Diagram: ?

Subunit composition of homoserine O-succinyltransferase = [MetA]2
         homoserine O-succinyltransferase = MetA

Summary:
Homoserine O-succinyltransferase (MetA) catalyzes the first unique step in the de novo methionine biosynthesis pathway in Escherichia coli (see pathway methionine biosynthesis I). It transfers a succinyl group from succinyl-CoA to homoserine. It is regulated at several levels and it appears to have a role in regulating bacterial growth. Its activity is regulated through allosteric feedback inhibition by the pathway end product methionine and also synergistically by the methionine derivative S-adenosylmethionine [Born99] (see pathway S-adenosyl-L-methionine cycle I). Expression of the enzyme is negatively regulated at the transcriptional level by the methionine regulon repressor MetJ and positively regulated by the activator MetR. MetA has also been shown to be a heat shock protein [Ron90, Biran95]. It was found to be very thermolabile, undergoing aggregation and proteolysis at elevated temperatures [Biran00, Gur02, Mordukhova08]. It appears to be regulated by temperature-dependent proteolysis, thereby mediating a change in growth rate as a function of temperature [Katz09].

Review: Hondrop, E.R. and R.G. Matthews (2006) "Methionine" EcoSal 3.6.1.7 [ECOSAL] .

The gene encoding this enzyme has been cloned, overexpressed, and its protein product characterized [Born99]. Site-directed mutagenesis and steady-state kinetic studies identified amino acid residues important to its catalytic mechanism [Rosen04, Ziegler07, Coe07] . Because this enzyme is not found in humans, it is of interest in the design of antimicrobials [Coe07]. The crystal structure of the orthologous MetA from Bacillus cereus in complex with homoserine has been determined at 2.0 Å resolution [Zubieta08].

The apparent molecular mass of the subunit was determined by SDS-PAGE and the apparent molecular mass of the native enzyme was determined by gel filtration chromatography [Born99].

Citations: [Duclos89]

Gene Citations: [Old93a]

Locations: cytosol

Map Position: [4,212,303 -> 4,213,232] (90.79 centisomes)
Length: 930 bp / 309 aa

Molecular Weight of Polypeptide: 35.727 kD (from nucleotide sequence), 35.6 kD (experimental) [Born99 ]

Molecular Weight of Multimer: 86.0 kD (experimental) [Born99]

Unification Links: ASAP:ABE-0013121 , CGSC:516 , EchoBASE:EB0576 , EcoGene:EG10581 , EcoliWiki:b4013 , Mint:MINT-1343536 , ModBase:P07623 , OU-Microarray:b4013 , PortEco:metA , PR:PRO_000023208 , Pride:P07623 , Protein Model Portal:P07623 , RefSeq:NP_418437 , RegulonDB:EG10581 , SMR:P07623 , String:511145.b4013 , Swiss-Model:P07623 , UniProt:P07623

Relationship Links: InterPro:IN-FAMILY:IPR005697 , Panther:IN-FAMILY:PTHR20919 , Pfam:IN-FAMILY:PF04204 , ProDom:IN-FAMILY:PD037892

Gene-Reaction Schematic: ?

Genetic Regulation Schematic: ?

GO Terms:

Biological Process: GO:0008152 - metabolic process Inferred by computational analysis [UniProtGOA11]
GO:0008652 - cellular amino acid biosynthetic process Inferred by computational analysis [UniProtGOA11]
GO:0009086 - methionine biosynthetic process Inferred by computational analysis [UniProtGOA11]
GO:0019281 - L-methionine biosynthetic process from homoserine via O-succinyl-L-homoserine and cystathionine Inferred by computational analysis [GOA06, GOA01a]
Molecular Function: GO:0008899 - homoserine O-succinyltransferase activity Inferred from experiment Inferred by computational analysis [GOA06, GOA01, GOA01a, Born99]
GO:0042803 - protein homodimerization activity Inferred from experiment [Born99]
GO:0003824 - catalytic activity Inferred by computational analysis [UniProtGOA11]
GO:0016740 - transferase activity Inferred by computational analysis [UniProtGOA11]
GO:0016746 - transferase activity, transferring acyl groups Inferred by computational analysis [UniProtGOA11]
Cellular Component: GO:0005737 - cytoplasm Inferred by computational analysis [UniProtGOA11a, UniProtGOA11, GOA06, GOA01a]
GO:0005829 - cytosol Inferred by computational analysis [DiazMejia09]

MultiFun Terms: metabolism biosynthesis of building blocks amino acids methionine

Essentiality data for metA 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 Indeterminate 37 Aerobic 7.2 0.22 Yes [Baba06, Comment 2]
No [Feist07, Comment 4]

Credits:
Last-Curated ? 22-Apr-2011 by Fulcher C , SRI International


Enzymatic reaction of: homoserine O-succinyltransferase

Synonyms: homoserine O-transsuccinylase, succinyl-CoA:L-homoserine O-succinyltransferase, homoserine succinyltransferase, O-succinylhomoserine synthetase, HTS

EC Number: 2.3.1.46

L-homoserine + succinyl-CoA <=> O-succinyl-L-homoserine + coenzyme A

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 favored in the direction shown.

Alternative Substrates for L-homoserine [Born99 ]: D-homoserine

In Pathways: superpathway of S-adenosyl-L-methionine biosynthesis , homoserine and methionine biosynthesis , superpathway of lysine, threonine and methionine biosynthesis I , aspartate superpathway , methionine biosynthesis I

Summary:
The enzyme employs a ping-pong mechanism in which the succinyl group of succinyl-CoA is first transferred to the enzyme, forming a succinyl-enzyme intermediate, and then transferred to homoserine to form the product O-succinylhomoserine. The equilibrium constant for the reaction strongly favored production of O-succinylhomoserine from succinyl-CoA [Born99].

Inhibitors (Allosteric): S-adenosyl-L-methionine [Comment 5] , L-methionine [Comment 5]

Inhibitors (Unknown Mechanism): diethylpyrocarbonate [Ziegler07] , iodoacetamide [Born99, Comment 6] , α-methylmethionine [Rowbury68]

Primary Physiological Regulators of Enzyme Activity: S-adenosyl-L-methionine , L-methionine

Kinetic Parameters:

Substrate
Km (μM)
kcat (sec-1)
kcat/Km (sec-1 μM-1)
Citations
succinyl-CoA
170.0
24.0
[Born99, BRENDA14]
succinyl-CoA
280.0
25.7
[Coe07, BRENDA14]
L-homoserine
720.0
[Ziegler07, BRENDA14]
L-homoserine
1600.0
[Born99, BRENDA14]
L-homoserine
380.0
[Coe07, BRENDA14]
O-succinyl-L-homoserine
3500.0
5.23
[Born99, BRENDA14]
coenzyme A
640.0
[Born99, BRENDA14]

pH(opt): 7.5 [BRENDA14, ROWBURY64]


Sequence Features

Feature Class Location Citations Comment
Cleavage-of-Initial-Methionine 1
[Born99, UniProt11]
UniProt: Removed.
Chain 2 -> 309
[UniProt09]
UniProt: Homoserine O-succinyltransferase;
Active-Site 47, 142, 235
[Ziegler07]
In the active site, the cysteine residue may act as a nucleophile in this reaction, and the histidine may serve to deprotonate the cysteine to allow that activity.
Sequence-Conflict 67
[Michaeli84, Duclos89, UniProt10a]
Alternate sequence: I → V; UniProt: (in Ref. 1 and 6);
Active-Site 142
[UniProt10]
UniProt: Non-Experimental Qualifier: potential;


Gene Local Context (not to scale): ?

Transcription Units:

Notes:

History:
10/20/97 Gene b4013 from Blattner lab Genbank (v. M52) entry merged into EcoCyc gene EG10581; 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

Biran00: Biran D, Gur E, Gollan L, Ron EZ (2000). "Control of methionine biosynthesis in Escherichia coli by proteolysis." Mol Microbiol 37(6);1436-43. PMID: 10998174

Biran95: Biran D, Brot N, Weissbach H, Ron EZ (1995). "Heat shock-dependent transcriptional activation of the metA gene of Escherichia coli." J Bacteriol 177(5);1374-9. PMID: 7868613

Born99: Born TL, Blanchard JS (1999). "Enzyme-catalyzed acylation of homoserine: mechanistic characterization of the Escherichia coli metA-encoded homoserine transsuccinylase." Biochemistry 1999;38(43);14416-23. PMID: 10572016

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

Coe07: Coe DM, Viola RE (2007). "Assessing the roles of essential functional groups in the mechanism of homoserine succinyltransferase." Arch Biochem Biophys 461(2);211-8. PMID: 17442255

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

Duclos89: Duclos B, Cortay JC, Bleicher F, Ron EZ, Richaud C, Saint Girons I, Cozzone AJ (1989). "Nucleotide sequence of the metA gene encoding homoserine trans-succinylase in Escherichia coli." Nucleic Acids Res 17(7);2856. PMID: 2654885

ECOSAL: "Escherichia coli and Salmonella: Cellular and Molecular Biology." Online edition.

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

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."

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

Gur02: Gur E, Biran D, Gazit E, Ron EZ (2002). "In vivo aggregation of a single enzyme limits growth of Escherichia coli at elevated temperatures." Mol Microbiol 46(5);1391-7. PMID: 12453224

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

Katz09: Katz C, Rasouly A, Gur E, Shenhar Y, Biran D, Ron EZ (2009). "Temperature-dependent proteolysis as a control element in Escherichia coli metabolism." Res Microbiol 160(9);684-6. PMID: 19770038

Lee66: Lee LW, Ravel JM, Shive W (1966). "Multimetabolite control of a biosynthetic pathway by sequential metabolites." J Biol Chem 1966;241(22);5479-80. PMID: 5333667

Michaeli84: Michaeli S, Mevarech M, Ron EZ (1984). "Regulatory region of the metA gene of Escherichia coli K-12." J Bacteriol 1984;160(3);1158-62. PMID: 6094503

Mordukhova08: Mordukhova EA, Lee HS, Pan JG (2008). "Improved thermostability and acetic acid tolerance of Escherichia coli via directed evolution of homoserine o-succinyltransferase." Appl Environ Microbiol 74(24);7660-8. PMID: 18978085

Old93a: Old IG, Saint Girons I, Richaud C (1993). "Physical mapping of the scattered methionine genes on the Escherichia coli chromosome." J Bacteriol 1993;175(11);3689-91. PMID: 8501076

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

Ron90: Ron EZ, Alajem S, Biran D, Grossman N (1990). "Adaptation of Escherichia coli to elevated temperatures: the metA gene product is a heat shock protein." Antonie Van Leeuwenhoek 1990;58(3);169-74. PMID: 2256677

Rosen04: Rosen R, Becher D, Buttner K, Biran D, Hecker M, Ron EZ (2004). "Probing the active site of homoserine trans-succinylase." FEBS Lett 577(3);386-92. PMID: 15556615

ROWBURY64: ROWBURY RJ, WOODS DD (1964). "O-SUCCINYLHOMOSERINE AS AN INTERMEDIATE IN THE SYNTHESIS OF CYSTATHIONINE BY ESCHERICHIA COLI." J Gen Microbiol 36;341-58. PMID: 14217349

Rowbury68: Rowbury RJ "The inhibitory action of alpha-methylmethionine on Escherichia coli." J Gen Microbiol 1968;52:223-230.

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

UniProt10: UniProt Consortium (2010). "UniProt version 2010-07 released on 2010-06-15 00:00:00." Database.

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

UniProt11: UniProt Consortium (2011). "UniProt version 2011-06 released on 2011-06-30 00:00:00." Database.

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."

Ziegler07: Ziegler K, Noble SM, Mutumanje E, Bishop B, Huddler DP, Born TL (2007). "Identification of catalytic cysteine, histidine, and lysine residues in Escherichia coli homoserine transsuccinylase." Biochemistry 46(10);2674-83. PMID: 17302437

Zubieta08: Zubieta C, Arkus KA, Cahoon RE, Jez JM (2008). "A single amino acid change is responsible for evolution of acyltransferase specificity in bacterial methionine biosynthesis." J Biol Chem 283(12);7561-7. PMID: 18216013

Other References Related to Gene Regulation

Gery97: Gery S, Ron EZ (1997). "An Escherichia coli gene divergently transcribed from a promoter overlapping the metA promoter." FEMS Microbiol Lett 154(2);219-22. PMID: 9311119

Liu01: Liu R, Blackwell TW, States DJ (2001). "Conformational model for binding site recognition by the E.coli MetJ transcription factor." Bioinformatics 17(7);622-33. PMID: 11448880

Marincs06: Marincs F, Manfield IW, Stead JA, McDowall KJ, Stockley PG (2006). "Transcript analysis reveals an extended regulon and the importance of protein-protein co-operativity for the Escherichia coli methionine repressor." Biochem J 396(2);227-34. PMID: 16515535


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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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