DB code: D00516

RLCP classification 3.113.90020.1182 : Transfer
3.1143.80000.1190 : Transfer
CATH domain 3.40.1190.10 : UDP-N-acetylmuramoyl-L-alanine Catalytic domain
3.90.190.20 : Protein-Tyrosine Phosphatase; Chain A
E.C. 6.3.2.17
CSA
M-CSA
MACiE

CATH domain Related DB codes (homologues)
3.40.1190.10 : UDP-N-acetylmuramoyl-L-alanine T00109
3.90.190.20 : Protein-Tyrosine Phosphatase; Chain A T00109

Uniprot Enzyme Name
UniprotKB Protein name Synonyms Pfam RefSeq
P15925 Folylpolyglutamate synthase
EC 6.3.2.17
Folylpoly-gamma-glutamate synthetase
FPGS
Tetrahydrofolylpolyglutamate synthase
Tetrahydrofolate synthase
PF02875 (Mur_ligase_C)
PF08245 (Mur_ligase_M)
[Graphical View]
Q9WY13
Folylpolyglutamate synthase/dihydrofolate synthase
PF02875 (Mur_ligase_C)
PF08245 (Mur_ligase_M)
[Graphical View] 		NP_227981.1 (Protein)
NC_000853.1 (DNA/RNA sequence)

KEGG enzyme name
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)

UniprotKB: Accession Number Entry name Activity Subunit Subcellular location Cofactor
P15925 FOLC_LACCA ATP + tetrahydropteroyl-(gamma-Glu)(n) + L- glutamate = ADP + phosphate + tetrahydropteroyl-(gamma-Glu)(n+1). Monomer.
Q9WY13 Q9WY13_THEMA

KEGG Pathways
Map code Pathways E.C.
MAP00790 Folate biosynthesis

Compound table
Cofactors Substrates Products Intermediates
KEGG-id C00305 C00002 C00025 C03541 C00008 C00009 C03541
E.C.
Compound Magnesium ATP L-Glutamate Tetrahydrofolyl-[Glu](n) ADP Orthophosphate Tetrahydrofolyl-[Glu](n+1)
Type divalent metal (Ca2+, Mg2+) amine group,nucleotide amino acids,carboxyl group amino acids,amide group,amine group,aromatic ring (only carbon atom),aromatic ring (with nitrogen atoms),carboxyl group,peptide/protein amine group,nucleotide phosphate group/phosphate ion amino acids,amide group,amine group,aromatic ring (only carbon atom),aromatic ring (with nitrogen atoms),carboxyl group,peptide/protein
ChEBI 18420
 18420
 		15422
 15422
 		16015
 16015
 		17420
 17420
 		16761
 16761
 		26078
 26078
 		17420
 17420
PubChem 888
 888
 		5957
 5957
 		33032
 44272391
 88747398
 33032
 44272391
 88747398
 		16722112
 442163
 16722112
 442163
 		6022
 6022
 		1004
 22486802
 1004
 22486802
 		16722112
 442163
 16722112
 442163
1fgsA01 Bound:_MG Unbound Unbound Unbound Unbound Analogue:POP Unbound
1jbvA01 Bound:2x_MG Analogue:ACP Unbound Unbound Unbound Unbound Unbound
1jbwA01 Bound:2x_MG Analogue:ACQ Unbound Analogue:TMF Unbound Unbound Unbound
1o5zA01 Unbound Unbound Unbound Unbound Unbound Unbound Unbound
1fgsA02 Unbound Unbound Unbound Unbound Unbound Unbound Unbound
1jbvA02 Unbound Unbound Unbound Unbound Unbound Unbound Unbound
1jbwA02 Unbound Unbound Unbound Unbound Unbound Unbound Unbound
1o5zA02 Unbound Unbound Unbound Unbound Unbound Unbound Unbound

Reference for Active-site residues
resource references E.C.

Active-site residues
PDB Catalytic residues Cofactor-binding residues Modified residues Main-chain involved in catalysis Comment
1fgsA01 LYS 50; SER 73;GLU 143(Magnesium-2); invisible 170-176
1jbvA01 LYS 50;HIS 170 SER 73;GLU 143(Magnesium-2);HIS 170(Magnesium-1)
1jbwA01 LYS 50;HIS 170 SER 73;GLU 143(Magnesium-2);HIS 170(Magnesium-1)
1o5zA01 LYS 51; SER 74;GLU 143(Magnesium-2); invisible 169-175
1fgsA02
1jbvA02
1jbwA02
1o5zA02

References for Catalytic Mechanism
References Sections No. 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]
Resource
Comments
Medline ID
PubMed ID 3838105
Journal Mol Pharmacol
Year 1985
Volume 27
Pages 156-66
Authors Moran RG, Colman PD, Rosowsky A, Forsch RA, Chan KK
Title Structural features of 4-amino antifolates required for substrate activity with mammalian folylpolyglutamate synthetase.
Related PDB
Related UniProtKB
[2]
Resource
Comments
Medline ID
PubMed ID 3828321
Journal Biochemistry
Year 1987
Volume 26
Pages 513-21
Authors Cichowicz DJ, Shane B
Title Mammalian folylpoly-gamma-glutamate synthetase. 2. Substrate specificity and kinetic properties.
Related PDB
Related UniProtKB
[3]
Resource
Comments
Medline ID
PubMed ID 3828322
Journal Biochemistry
Year 1987
Volume 26
Pages 522-9
Authors George S, Cichowicz DJ, Shane B
Title Mammalian folylpoly-gamma-glutamate synthetase. 3. Specificity for folate analogues.
Related PDB
Related UniProtKB
[4]
Resource
Comments
Medline ID
PubMed ID 2906805
Journal Biochemistry
Year 1988
Volume 27
Pages 9062-70
Authors Banerjee RV, Shane B, McGuire JJ, Coward JK
Title Dihydrofolate synthetase and folylpolyglutamate synthetase: direct evidence for intervention of acyl phosphate intermediates.
Related PDB
Related UniProtKB
[5]
Resource
Comments
Medline ID
PubMed ID 2168155
Journal Arch Biochem Biophys
Year 1990
Volume 281
Pages 198-203
Authors Bolanowska WE, Russell CA, McGuire JJ
Title Activation of mammalian folylpolyglutamate synthetase by sodium bicarbonate.
Related PDB
Related UniProtKB
[6]
Resource
Comments
Medline ID
PubMed ID 1989505
Journal Arch Biochem Biophys
Year 1991
Volume 284
Pages 9-16
Authors Kimlova LJ, Pyne C, Keshavjee K, Huy J, Beebakhee G, Bognar AL
Title Mutagenesis of the folC gene encoding folylpolyglutamate synthetase-dihydrofolate synthetase in Escherichia coli.
Related PDB
Related UniProtKB
[7]
Resource
Comments
Medline ID
PubMed ID 1578484
Journal J Med Chem
Year 1992
Volume 35
Pages 1578-88
Authors Rosowsky A, Forsch RA, Reich VE, Freisheim JH, Moran RG
Title Side chain modified 5-deazafolate and 5-deazatetrahydrofolate analogues as mammalian folylpolyglutamate synthetase and glycinamide ribonucleotide formyltransferase inhibitors: synthesis and in vitro biological evaluation.
Related PDB
Related UniProtKB
[8]
Resource
Comments
Medline ID
PubMed ID 1375963
Journal J Med Chem
Year 1992
Volume 35
Pages 2002-6
Authors Singh SK, Singer SC, Ferone R, Waters KA, Mullin RJ, Hynes JB
Title Synthesis and biological evaluation of N alpha-(5-deaza-5,6,7,8-tetrahydropteroyl)-L-ornithine.
Related PDB
Related UniProtKB
[9]
Resource
Comments
Medline ID
PubMed ID 1569575
Journal J Mol Biol
Year 1992
Volume 224
Pages 1179-80
Authors Cody V, Luft JR, Pangborn W, Toy J, Bognar AL
Title Purification and crystallization of Lactobacillus casei folylpolyglutamate synthetase expressed in Escherichia coli.
Related PDB
Related UniProtKB
[10]
Resource
Comments
Medline ID
PubMed ID 7979375
Journal Arch Biochem Biophys
Year 1994
Volume 314
Pages 344-50
Authors Toy J, Bognar AL
Title Mutagenesis of the Lactobacillus casei folylpolyglutamate synthetase gene at essential residues resembling an ATP binding site.
Related PDB
Related UniProtKB
[11]
Resource
Comments
Medline ID
PubMed ID 8114682
Journal Mol Pharmacol
Year 1994
Volume 45
Pages 341-51
Authors Sanghani PC, Jackman A, Evans VR, Thornton T, Hughes L, Calvert AH, Moran RG
Title A strategy for the design of membrane-permeable folypoly-gamma-glutamate synthetase inhibitors: "bay-region"-substituted 2-desamino-2-methyl-5,8-dideazafolate analogs.
Related PDB
Related UniProtKB
[12]
Resource
Comments
Medline ID
PubMed ID 9166795
Journal Biochemistry
Year 1997
Volume 36
Pages 6223-9
Authors Eveland SS, Pompliano DL, Anderson MS
Title Conditionally 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.
Related PDB
Related UniProtKB
[13]
Resource
Comments X-RAY CRYSTALLOGRAPHY (2.4 ANGSTROMS).
Medline ID 98283985
PubMed ID 9618466
Journal Proc Natl Acad Sci U S A
Year 1998
Volume 95
Pages 6647-52
Authors Sun X, Bognar AL, Baker EN, Smith CA
Title Structural homologies with ATP- and folate-binding enzymes in the crystal structure of folylpolyglutamate synthetase.
Related PDB 1fgs
Related UniProtKB P15925
[14]
Resource
Comments
Medline ID
PubMed ID 10479284
Journal J Med Chem
Year 1999
Volume 42
Pages 3510-9
Authors Rosowsky A, Forsch RA, Null A, Moran RG
Title 5-deazafolate analogues with a rotationally restricted glutamate or ornithine side chain: synthesis and binding interaction with folylpolyglutamate synthetase.
Related PDB
Related UniProtKB
[15]
Resource
Comments
Medline ID
PubMed ID 10966819
Journal J Mol Biol
Year 2000
Volume 301
Pages 1257-66
Authors Bertrand 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.
Related PDB
Related UniProtKB
[16]
Resource
Comments
Medline ID
PubMed ID 10970743
Journal J Mol Biol
Year 2000
Volume 302
Pages 427-40
Authors Sheng Y, Sun X, Shen Y, Bognar AL, Baker EN, Smith CA
Title Structural and functional similarities in the ADP-forming amide bond ligase superfamily: implications for a substrate-induced conformational change in folylpolyglutamate synthetase.
Related PDB
Related UniProtKB
[17]
Resource
Comments X-ray crystallography
Medline ID
PubMed ID 11501996
Journal J Mol Biol
Year 2001
Volume 310
Pages 1067-78
Authors Sun X, Cross JA, Bognar AL, Baker EN, Smith CA
Title Folate-binding triggers the activation of folylpolyglutamate synthetase.
Related PDB 1jbv 1jbw
Related UniProtKB
[18]
Resource
Comments
Medline ID
PubMed ID 12051687
Journal Arch Biochem Biophys
Year 2002
Volume 402
Pages 94-103
Authors Sheng Y, Cross JA, Shen Y, Smith CA, Bognar AL
Title Mutation of an essential glutamate residue in folylpolyglutamate synthetase and activation of the enzyme by pteroate binding.
Related PDB
Related UniProtKB
[19]
Resource
Comments
Medline ID
PubMed ID 12578366
Journal Biochemistry
Year 2003
Volume 42
Pages 1537-43
Authors Sheng Y, Ip H, Liu J, Davidson A, Bognar AL
Title Binding of ATP as well as tetrahydrofolate induces conformational changes in Lactobacillus casei folylpolyglutamate synthetase in solution.
Related PDB
Related UniProtKB

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.

Created Updated
2004-03-25 2009-02-26