EzCatDB: D00516
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DB codeD00516
RLCP classification3.113.90020.1182 : Transfer
3.1143.80000.1190 : Transfer
CATH domainDomain 13.40.1190.10 : UDP-N-acetylmuramoyl-L-alanineCatalytic domain
Domain 23.90.190.20 : Protein-Tyrosine Phosphatase; Chain A
E.C.6.3.2.17

CATH domainRelated DB codes (homologues)
3.40.1190.10 : UDP-N-acetylmuramoyl-L-alanineT00109
3.90.190.20 : Protein-Tyrosine Phosphatase; Chain AT00109

Enzyme Name
UniProtKBKEGG

P15925Q9WY13
Protein nameFolylpolyglutamate synthase
tetrahydrofolate synthase
folylpolyglutamate synthase
folate polyglutamate synthetase
formyltetrahydropteroyldiglutamate synthetase
N10-formyltetrahydropteroyldiglutamate synthetase
folylpoly-gamma-glutamate synthase
folylpolyglutamyl synthetase
folylpoly(gamma-glutamate) synthase
folylpolyglutamate synthetase
folylpoly-gamma-glutamate synthetase-dihydrofolate synthetase
FPGS
tetrahydrofolylpolyglutamate synthase
tetrahydrofolate:L-glutamate gamma-ligase (ADP-forming)
tetrahydropteroyl-[gamma-Glu]n:L-glutamate gamma-ligase(ADP-forming)
SynonymsEC 6.3.2.17
Folylpoly-gamma-glutamate synthetase
FPGS
Tetrahydrofolylpolyglutamate synthase
Tetrahydrofolate synthase
Folylpolyglutamate synthase/dihydrofolate synthase
RefSeq
NP_227981.1 (Protein)
NC_000853.1 (DNA/RNA sequence)
PfamPF02875 (Mur_ligase_C)
PF08245 (Mur_ligase_M)
[Graphical view]
PF02875 (Mur_ligase_C)
PF08245 (Mur_ligase_M)
[Graphical view]

KEGG pathways
MAP codePathways
MAP00790Folate biosynthesis

UniProtKB:Accession NumberP15925Q9WY13
Entry nameFOLC_LACCAQ9WY13_THEMA
ActivityATP + tetrahydropteroyl-(gamma-Glu)(n) + L- glutamate = ADP + phosphate + tetrahydropteroyl-(gamma-Glu)(n+1).
SubunitMonomer.
Subcellular location

Cofactor


Compound table: links to PDB-related databases & PoSSuM

CofactorsSubstratesProducts
KEGG-idC00305C00002C00025C03541C00008C00009C03541
CompoundMagnesiumATPL-GlutamateTetrahydrofolyl-[Glu](n)ADPOrthophosphateTetrahydrofolyl-[Glu](n+1)
Typedivalent metal (Ca2+, Mg2+)amine group,nucleotideamino acids,carboxyl groupamino acids,amide group,amine group,aromatic ring (only carbon atom),aromatic ring (with nitrogen atoms),carboxyl group,peptide/proteinamine group,nucleotidephosphate group/phosphate ionamino acids,amide group,amine group,aromatic ring (only carbon atom),aromatic ring (with nitrogen atoms),carboxyl group,peptide/protein
ChEBI18420
15422
16015
17420
16761
26078
17420
PubChem888
5957
88747398
44272391
33032
442163
16722112
6022
22486802
1004
442163
16722112
               
1fgsA01Bound:_MGUnboundUnboundUnboundUnboundAnalogue:POPUnbound
1jbvA01Bound:2x_MGAnalogue:ACPUnboundUnboundUnboundUnboundUnbound
1jbwA01Bound:2x_MGAnalogue:ACQUnboundAnalogue:TMFUnboundUnboundUnbound
1o5zA01UnboundUnboundUnboundUnboundUnboundUnboundUnbound
1fgsA02UnboundUnboundUnboundUnboundUnboundUnboundUnbound
1jbvA02UnboundUnboundUnboundUnboundUnboundUnboundUnbound
1jbwA02UnboundUnboundUnboundUnboundUnboundUnboundUnbound
1o5zA02UnboundUnboundUnboundUnboundUnboundUnboundUnbound

Active-site residues
pdbCatalytic residuesCofactor-binding residuescomment
           
1fgsA01LYS 50;       
SER 73;GLU 143(Magnesium-2);                    
invisible 170-176
1jbvA01LYS 50;HIS 170
SER 73;GLU 143(Magnesium-2);HIS 170(Magnesium-1)
 
1jbwA01LYS 50;HIS 170
SER 73;GLU 143(Magnesium-2);HIS 170(Magnesium-1)
 
1o5zA01LYS 51;       
SER 74;GLU 143(Magnesium-2);                    
invisible 169-175
1fgsA02 
 
 
1jbvA02 
 
 
1jbwA02 
 
 
1o5zA02 
 
 

References for Catalytic Mechanism
ReferencesSectionsNo. of steps in catalysis
[3]p.527-528
[4]

[12]Fig.2A
[15]p.1263-1264
[16]Fig.1c
[17]p.1069-1074

references
[1]
PubMed ID3838105
JournalMol Pharmacol
Year1985
Volume27
Pages156-66
AuthorsMoran RG, Colman PD, Rosowsky A, Forsch RA, Chan KK
TitleStructural features of 4-amino antifolates required for substrate activity with mammalian folylpolyglutamate synthetase.
[2]
PubMed ID3828321
JournalBiochemistry
Year1987
Volume26
Pages513-21
AuthorsCichowicz DJ, Shane B
TitleMammalian folylpoly-gamma-glutamate synthetase. 2. Substrate specificity and kinetic properties.
[3]
PubMed ID3828322
JournalBiochemistry
Year1987
Volume26
Pages522-9
AuthorsGeorge S, Cichowicz DJ, Shane B
TitleMammalian folylpoly-gamma-glutamate synthetase. 3. Specificity for folate analogues.
[4]
PubMed ID2906805
JournalBiochemistry
Year1988
Volume27
Pages9062-70
AuthorsBanerjee RV, Shane B, McGuire JJ, Coward JK
TitleDihydrofolate synthetase and folylpolyglutamate synthetase: direct evidence for intervention of acyl phosphate intermediates.
[5]
PubMed ID2168155
JournalArch Biochem Biophys
Year1990
Volume281
Pages198-203
AuthorsBolanowska WE, Russell CA, McGuire JJ
TitleActivation of mammalian folylpolyglutamate synthetase by sodium bicarbonate.
[6]
PubMed ID1989505
JournalArch Biochem Biophys
Year1991
Volume284
Pages9-16
AuthorsKimlova LJ, Pyne C, Keshavjee K, Huy J, Beebakhee G, Bognar AL
TitleMutagenesis of the folC gene encoding folylpolyglutamate synthetase-dihydrofolate synthetase in Escherichia coli.
[7]
PubMed ID1578484
JournalJ Med Chem
Year1992
Volume35
Pages1578-88
AuthorsRosowsky A, Forsch RA, Reich VE, Freisheim JH, Moran RG
TitleSide chain modified 5-deazafolate and 5-deazatetrahydrofolate analogues as mammalian folylpolyglutamate synthetase and glycinamide ribonucleotide formyltransferase inhibitors: synthesis and in vitro biological evaluation.
[8]
PubMed ID1375963
JournalJ Med Chem
Year1992
Volume35
Pages2002-6
AuthorsSingh SK, Singer SC, Ferone R, Waters KA, Mullin RJ, Hynes JB
TitleSynthesis and biological evaluation of N alpha-(5-deaza-5,6,7,8-tetrahydropteroyl)-L-ornithine.
[9]
PubMed ID1569575
JournalJ Mol Biol
Year1992
Volume224
Pages1179-80
AuthorsCody V, Luft JR, Pangborn W, Toy J, Bognar AL
TitlePurification and crystallization of Lactobacillus casei folylpolyglutamate synthetase expressed in Escherichia coli.
[10]
PubMed ID7979375
JournalArch Biochem Biophys
Year1994
Volume314
Pages344-50
AuthorsToy J, Bognar AL
TitleMutagenesis of the Lactobacillus casei folylpolyglutamate synthetase gene at essential residues resembling an ATP binding site.
[11]
PubMed ID8114682
JournalMol Pharmacol
Year1994
Volume45
Pages341-51
AuthorsSanghani PC, Jackman A, Evans VR, Thornton T, Hughes L, Calvert AH, Moran RG
TitleA strategy for the design of membrane-permeable folypoly-gamma-glutamate synthetase inhibitors: "bay-region"-substituted 2-desamino-2-methyl-5,8-dideazafolate analogs.
[12]
PubMed ID9166795
JournalBiochemistry
Year1997
Volume36
Pages6223-9
AuthorsEveland SS, Pompliano DL, Anderson MS
TitleConditionally lethal Escherichia coli murein mutants contain point defects that map to regions conserved among murein and folyl poly-gamma-glutamate ligases: identification of a ligase superfamily.
[13]
CommentsX-RAY CRYSTALLOGRAPHY (2.4 ANGSTROMS).
Medline ID98283985
PubMed ID9618466
JournalProc Natl Acad Sci U S A
Year1998
Volume95
Pages6647-52
AuthorsSun X, Bognar AL, Baker EN, Smith CA
TitleStructural homologies with ATP- and folate-binding enzymes in the crystal structure of folylpolyglutamate synthetase.
Related PDB1fgs
Related UniProtKBP15925
[14]
PubMed ID10479284
JournalJ Med Chem
Year1999
Volume42
Pages3510-9
AuthorsRosowsky A, Forsch RA, Null A, Moran RG
Title5-deazafolate analogues with a rotationally restricted glutamate or ornithine side chain: synthesis and binding interaction with folylpolyglutamate synthetase.
[15]
PubMed ID10966819
JournalJ Mol Biol
Year2000
Volume301
Pages1257-66
AuthorsBertrand JA, Fanchon E, Martin L, Chantalat L, Auger G, Blanot D, van Heijenoort J, Dideberg O
Title"Open" structures of MurD: domain movements and structural similarities with folylpolyglutamate synthetase.
[16]
PubMed ID10970743
JournalJ Mol Biol
Year2000
Volume302
Pages427-40
AuthorsSheng Y, Sun X, Shen Y, Bognar AL, Baker EN, Smith CA
TitleStructural and functional similarities in the ADP-forming amide bond ligase superfamily: implications for a substrate-induced conformational change in folylpolyglutamate synthetase.
[17]
CommentsX-ray crystallography
PubMed ID11501996
JournalJ Mol Biol
Year2001
Volume310
Pages1067-78
AuthorsSun X, Cross JA, Bognar AL, Baker EN, Smith CA
TitleFolate-binding triggers the activation of folylpolyglutamate synthetase.
Related PDB1jbv,1jbw
[18]
PubMed ID12051687
JournalArch Biochem Biophys
Year2002
Volume402
Pages94-103
AuthorsSheng Y, Cross JA, Shen Y, Smith CA, Bognar AL
TitleMutation of an essential glutamate residue in folylpolyglutamate synthetase and activation of the enzyme by pteroate binding.
[19]
PubMed ID12578366
JournalBiochemistry
Year2003
Volume42
Pages1537-43
AuthorsSheng Y, Ip H, Liu J, Davidson A, Bognar AL
TitleBinding of ATP as well as tetrahydrofolate induces conformational changes in Lactobacillus casei folylpolyglutamate synthetase in solution.

comments
According to the literature [15] & [17], this enzyme adopts a similar catalytic mechanism to that of UDP-N-acetylmuramoyl-L-alanine, as they are homologous with a similar catalytic site (T00109 in EzCatDB).
Firstly, it transfers phosphate group from ATP to the carboxyl group of the second substrate, tetrahydrofolyl-[Glu](n) (THF-[Glu]n), forming an acyl-phosphate intermediate. Secondly, it transfers acyl group from the intermediate to the amine group of the third substrate, L-glutamate (L-Glu), releasing the inorganic phosphate.
The first reaction (phosphate transfer) occurs as follows (see [15] & [17]):
(1) The acceptor group, carboxyl oxygen atom of THF-[Glu]n, makes a nucleophilic attack on the transferred group, the phosphorus atom of the gamma-phosphate group of ATP. (The reaction prpbably proceeds through an SN2-mechanism.)
(2) Lys50 and Mg2+ at site-2 stabilize the transition-state by neutralizing the transferred and leaving group, which are the beta- and gamma-phosphate groups of ATP, respectively.
(3) Mg2+ at site-1 activates the acceptor group, the carboxyl group of UMA, and stabilizes the transferred group, the gamma-phosphate group of ATP.
The second reaction (acyl transfer) occurs as follows (see [17] & T00109 in EzCatDB):
(1') The first base, probably the gamma-phosphate group of the substrate, may abstract the proton from the second acceptor group, the amine group of L-Glu.
(2') After the third substrate, L-Glu, was bound to the active site, the acceptor group, the amine group of L-Glu, would make a nucleophilic attack on the second transferred group, the carbonyl carbon of phosphorylated-THF-[Glu]n, forming the tetrahedral carbon intermediate.
(3') The second base, probably His170, may abstract the proton from the amine of the tetrahedral intermediate, facilitating the transformation of the intermediate into the final product and the release of the inorganic phosphate.

createdupdated
2004-03-252009-02-26


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