FPGS_LACCA
ID FPGS_LACCA Reviewed; 428 AA.
AC P15925;
DT 01-APR-1990, integrated into UniProtKB/Swiss-Prot.
DT 01-APR-1990, sequence version 1.
DT 03-AUG-2022, entry version 119.
DE RecName: Full=Folylpolyglutamate synthase {ECO:0000305};
DE Short=FPGS {ECO:0000303|PubMed:11501996};
DE EC=6.3.2.17 {ECO:0000269|PubMed:6138353};
DE AltName: Full=Folylpoly-gamma-glutamate synthetase {ECO:0000303|PubMed:2105929};
DE AltName: Full=Tetrahydrofolylpolyglutamate synthase;
GN Name=fpgS {ECO:0000312|EMBL:AAA88210.1};
OS Lactobacillus casei.
OC Bacteria; Firmicutes; Bacilli; Lactobacillales; Lactobacillaceae;
OC Lacticaseibacillus.
OX NCBI_TaxID=1582;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=ATCC 7469;
RX PubMed=2105929; DOI=10.1016/s0021-9258(19)39827-8;
RA Toy J., Bognar A.L.;
RT "Cloning and expression of the gene encoding Lactobacillus casei folylpoly-
RT gamma-glutamate synthetase in Escherichia coli and determination of its
RT primary structure.";
RL J. Biol. Chem. 265:2492-2499(1990).
RN [2]
RP FUNCTION, CATALYTIC ACTIVITY, SUBSTRATE SPECIFICITY, BIOPHYSICOCHEMICAL
RP PROPERTIES, COFACTOR, ACTIVITY REGULATION, SUBUNIT, AND REACTION MECHANISM.
RX PubMed=6138353; DOI=10.1016/s0021-9258(17)44215-3;
RA Bognar A.L., Shane B.;
RT "Purification and properties of Lactobacillus casei folylpoly-gamma-
RT glutamate synthetase.";
RL J. Biol. Chem. 258:12574-12581(1983).
RN [3]
RP FUNCTION, MUTAGENESIS OF ASP-151, CHIMERA PROTEINS, CATALYTIC ACTIVITY, AND
RP BIOPHYSICOCHEMICAL PROPERTIES.
RX PubMed=18232714; DOI=10.1021/bi701670y;
RA Sheng Y., Khanam N., Tsaksis Y., Shi X.M., Lu Q.S., Bognar A.L.;
RT "Mutagenesis of folylpolyglutamate synthetase indicates that
RT dihydropteroate and tetrahydrofolate bind to the same site.";
RL Biochemistry 47:2388-2396(2008).
RN [4]
RP X-RAY CRYSTALLOGRAPHY (2.4 ANGSTROMS).
RX PubMed=9618466; DOI=10.1073/pnas.95.12.6647;
RA Sun X., Bognar A.L., Baker E.N., Smith C.A.;
RT "Structural homologies with ATP- and folate-binding enzymes in the crystal
RT structure of folylpolyglutamate synthetase.";
RL Proc. Natl. Acad. Sci. U.S.A. 95:6647-6652(1998).
RN [5]
RP X-RAY CRYSTALLOGRAPHY (1.85 ANGSTROMS) IN COMPLEXES WITH ATP ANALOG;
RP MAGNESIUM AND 5,10-METHYLENETETRAHYDROFOLATE, COFACTOR, CARBOXYLATION AT
RP LYS-185, AND MUTAGENESIS OF HIS-316 AND SER-412.
RX PubMed=11501996; DOI=10.1006/jmbi.2001.4815;
RA Sun X., Cross J.A., Bognar A.L., Baker E.N., Smith C.A.;
RT "Folate-binding triggers the activation of folylpolyglutamate synthetase.";
RL J. Mol. Biol. 310:1067-1078(2001).
RN [6]
RP X-RAY CRYSTALLOGRAPHY (1.85 ANGSTROMS) OF WILD-TYPE AND MUTANTS SER-51;
RP 51-SER-THR-52 AND ALA-73.
RX PubMed=16627949; DOI=10.1107/s0907444906009796;
RA Smith C.A., Cross J.A., Bognar A.L., Sun X.;
RT "Mutation of Gly51 to serine in the P-loop of Lactobacillus casei
RT folylpolyglutamate synthetase abolishes activity by altering the
RT conformation of two adjacent loops.";
RL Acta Crystallogr. D 62:548-558(2006).
CC -!- FUNCTION: Involved in the conversion of folates to polyglutamate
CC derivatives, and likely functions in the retention of cellular folate
CC pools. Catalyzes successive MgATP-dependent additions of glutamate to a
CC pteroylmonoglutamate substrate, with a high preference for 5,10-
CC methylenetetrahydrofolate (mTHF). Thus, metabolizes mTHF to the
CC tetraglutamate derivative, but longer glutamate chain length products
CC are not observed. Tetrahydrofolate (H4PteGlu) and 10-formyl-H4PteGlu
CC are poorer folate substrates. In contrast to E.coli FolC, this enzyme
CC does not display dihydrofolate synthase activity.
CC {ECO:0000269|PubMed:18232714, ECO:0000269|PubMed:6138353}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=(6S)-5,6,7,8-tetrahydrofolyl-(gamma-L-Glu)(n) + ATP + L-
CC glutamate = (6S)-5,6,7,8-tetrahydrofolyl-(gamma-L-Glu)(n+1) + ADP +
CC H(+) + phosphate; Xref=Rhea:RHEA:10580, Rhea:RHEA-COMP:14738,
CC Rhea:RHEA-COMP:14740, ChEBI:CHEBI:15378, ChEBI:CHEBI:29985,
CC ChEBI:CHEBI:30616, ChEBI:CHEBI:43474, ChEBI:CHEBI:141005,
CC ChEBI:CHEBI:456216; EC=6.3.2.17;
CC Evidence={ECO:0000269|PubMed:6138353};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=(6R)-5,10-methylenetetrahydrofolyl-(gamma-L-Glu)(n) + ATP + L-
CC glutamate = (6R)-5,10-methylenetetrahydrofolyl-(gamma-L-Glu)(n+1) +
CC ADP + H(+) + phosphate; Xref=Rhea:RHEA:51912, Rhea:RHEA-COMP:13257,
CC Rhea:RHEA-COMP:13258, ChEBI:CHEBI:15378, ChEBI:CHEBI:29985,
CC ChEBI:CHEBI:30616, ChEBI:CHEBI:43474, ChEBI:CHEBI:136572,
CC ChEBI:CHEBI:456216; EC=6.3.2.17;
CC Evidence={ECO:0000269|PubMed:18232714, ECO:0000269|PubMed:6138353};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=10-formyltetrahydrofolyl-(gamma-L-Glu)(n) + ATP + L-glutamate
CC = 10-formyltetrahydrofolyl-(gamma-L-Glu)(n+1) + ADP + H(+) +
CC phosphate; Xref=Rhea:RHEA:51904, Rhea:RHEA-COMP:13088, Rhea:RHEA-
CC COMP:14300, ChEBI:CHEBI:15378, ChEBI:CHEBI:29985, ChEBI:CHEBI:30616,
CC ChEBI:CHEBI:43474, ChEBI:CHEBI:134413, ChEBI:CHEBI:456216;
CC EC=6.3.2.17; Evidence={ECO:0000269|PubMed:6138353};
CC -!- COFACTOR:
CC Name=Mg(2+); Xref=ChEBI:CHEBI:18420;
CC Evidence={ECO:0000269|PubMed:6138353};
CC Note=Binds 2 Mg(2+) ions per subunit. {ECO:0000269|PubMed:11501996};
CC -!- ACTIVITY REGULATION: Competitively inhibited by adenosine 5'-(3-
CC thio)triphosphate and beta,gamma-methylene-ATP.
CC {ECO:0000269|PubMed:6138353}.
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=2.3 uM for (6R)-5,10-methylenetetrahydropteroyldiglutamate
CC {ECO:0000269|PubMed:6138353};
CC KM=5.6 mM for ATP {ECO:0000269|PubMed:6138353};
CC KM=423 uM for glutamate {ECO:0000269|PubMed:6138353};
CC KM=32 uM for (6RS)-5,10-methylenetetrahydrofolate
CC {ECO:0000269|PubMed:18232714};
CC KM=3.4 mM for ATP {ECO:0000269|PubMed:18232714};
CC KM=470 uM for glutamate {ECO:0000269|PubMed:18232714};
CC KM=27 uM for (6RS)-5,10-methylenetetrahydropteroyldiglutamate
CC {ECO:0000269|PubMed:18232714};
CC Vmax=67.5 umol/h/mg enzyme with (6R)-5,10-
CC methylenetetrahydropteroyldiglutamate as substrate
CC {ECO:0000269|PubMed:6138353};
CC Vmax=19 umol/h/mg enzyme with (6RS)-5,10-methylenetetrahydrofolate as
CC substrate {ECO:0000269|PubMed:18232714};
CC Vmax=14 umol/h/mg enzyme with (6RS)-5,10-
CC methylenetetrahydropteroyldiglutamate as substrate
CC {ECO:0000269|PubMed:18232714};
CC pH dependence:
CC Optimum pH is about 10. {ECO:0000269|PubMed:6138353};
CC -!- SUBUNIT: Monomer. {ECO:0000269|PubMed:6138353}.
CC -!- MISCELLANEOUS: In contrast to many bacteria such as E.coli and
CC Corynebacterium spp., L.casei cannot synthesize folate de novo, and
CC requires exogenous folates for growth. L.casei metabolizes folate to
CC polyglutamates of chain length up to 11, with octa- and nonaglutamates
CC predominating. {ECO:0000305|PubMed:6138353}.
CC -!- MISCELLANEOUS: Kinetic studies are consistent with an ordered Ter-Ter
CC mechanism with MgATP binding first to the enzyme, folate second, and
CC glutamate last. The order of product dissociation from the enzyme is
CC ADP, folate product, and Pi. {ECO:0000269|PubMed:6138353}.
CC -!- SIMILARITY: Belongs to the folylpolyglutamate synthase family.
CC {ECO:0000305}.
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DR EMBL; J05221; AAA88210.1; -; Genomic_DNA.
DR PIR; A35534; A35534.
DR PDB; 1FGS; X-ray; 2.40 A; A=1-428.
DR PDB; 1JBV; X-ray; 1.95 A; A=1-428.
DR PDB; 1JBW; X-ray; 1.85 A; A=1-428.
DR PDB; 2GC5; X-ray; 1.85 A; A=1-428.
DR PDB; 2GC6; X-ray; 1.90 A; A=1-428.
DR PDB; 2GCA; X-ray; 2.40 A; A=1-428.
DR PDB; 2GCB; X-ray; 2.30 A; A=1-428.
DR PDBsum; 1FGS; -.
DR PDBsum; 1JBV; -.
DR PDBsum; 1JBW; -.
DR PDBsum; 2GC5; -.
DR PDBsum; 2GC6; -.
DR PDBsum; 2GCA; -.
DR PDBsum; 2GCB; -.
DR AlphaFoldDB; P15925; -.
DR SMR; P15925; -.
DR DrugBank; DB02301; 5,10-Methylene-6-Hydrofolic Acid.
DR DrugBank; DB03755; Adenosine-5'-[Beta, Gamma-Methylene]Tetraphosphate.
DR DrugBank; DB03909; Adenosine-5'-[Beta, Gamma-Methylene]Triphosphate.
DR DrugBank; DB03801; Lysine Nz-Carboxylic Acid.
DR BRENDA; 6.3.2.17; 2854.
DR SABIO-RK; P15925; -.
DR EvolutionaryTrace; P15925; -.
DR GO; GO:0005524; F:ATP binding; IEA:UniProtKB-KW.
DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
DR GO; GO:0004326; F:tetrahydrofolylpolyglutamate synthase activity; IEA:UniProtKB-EC.
DR GO; GO:0006730; P:one-carbon metabolic process; IEA:UniProtKB-KW.
DR Gene3D; 3.40.1190.10; -; 1.
DR Gene3D; 3.90.190.20; -; 1.
DR InterPro; IPR001645; Folylpolyglutamate_synth.
DR InterPro; IPR018109; Folylpolyglutamate_synth_CS.
DR InterPro; IPR036565; Mur-like_cat_sf.
DR InterPro; IPR004101; Mur_ligase_C.
DR InterPro; IPR036615; Mur_ligase_C_dom_sf.
DR InterPro; IPR013221; Mur_ligase_cen.
DR PANTHER; PTHR11136; PTHR11136; 1.
DR Pfam; PF02875; Mur_ligase_C; 1.
DR Pfam; PF08245; Mur_ligase_M; 1.
DR PIRSF; PIRSF001563; Folylpolyglu_synth; 1.
DR SUPFAM; SSF53244; SSF53244; 1.
DR SUPFAM; SSF53623; SSF53623; 1.
DR TIGRFAMs; TIGR01499; folC; 1.
DR PROSITE; PS01011; FOLYLPOLYGLU_SYNT_1; 1.
DR PROSITE; PS01012; FOLYLPOLYGLU_SYNT_2; 1.
PE 1: Evidence at protein level;
KW 3D-structure; ATP-binding; Ligase; Magnesium; Metal-binding;
KW Nucleotide-binding; One-carbon metabolism.
FT CHAIN 1..428
FT /note="Folylpolyglutamate synthase"
FT /id="PRO_0000168305"
FT BINDING 49..52
FT /ligand="ATP"
FT /ligand_id="ChEBI:CHEBI:30616"
FT /evidence="ECO:0000305|PubMed:11501996,
FT ECO:0007744|PDB:1JBV, ECO:0007744|PDB:1JBW"
FT BINDING 73
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="1"
FT /evidence="ECO:0000269|PubMed:11501996,
FT ECO:0007744|PDB:1JBV, ECO:0007744|PDB:1JBW"
FT BINDING 75
FT /ligand="(6R)-5,10-methylenetetrahydrofolyl-(gamma-L-Glu)n"
FT /ligand_id="ChEBI:CHEBI:136572"
FT /evidence="ECO:0000305|PubMed:11501996,
FT ECO:0007744|PDB:1JBW"
FT BINDING 82
FT /ligand="(6R)-5,10-methylenetetrahydrofolyl-(gamma-L-Glu)n"
FT /ligand_id="ChEBI:CHEBI:136572"
FT /evidence="ECO:0000305|PubMed:11501996,
FT ECO:0007744|PDB:1JBW"
FT BINDING 143
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="1"
FT /evidence="ECO:0000269|PubMed:11501996,
FT ECO:0007744|PDB:1JBV, ECO:0007744|PDB:1JBW"
FT BINDING 170
FT /ligand="Mg(2+)"
FT /ligand_id="ChEBI:CHEBI:18420"
FT /ligand_label="2"
FT /evidence="ECO:0000269|PubMed:11501996,
FT ECO:0007744|PDB:1JBV"
FT BINDING 264
FT /ligand="ATP"
FT /ligand_id="ChEBI:CHEBI:30616"
FT /evidence="ECO:0000305|PubMed:11501996,
FT ECO:0007744|PDB:1JBW"
FT BINDING 300
FT /ligand="ATP"
FT /ligand_id="ChEBI:CHEBI:30616"
FT /evidence="ECO:0000305|PubMed:11501996,
FT ECO:0007744|PDB:1JBV, ECO:0007744|PDB:1JBW"
FT BINDING 313..316
FT /ligand="ATP"
FT /ligand_id="ChEBI:CHEBI:30616"
FT /evidence="ECO:0000305|PubMed:11501996,
FT ECO:0007744|PDB:1JBV, ECO:0007744|PDB:1JBW"
FT BINDING 417
FT /ligand="(6R)-5,10-methylenetetrahydrofolyl-(gamma-L-Glu)n"
FT /ligand_id="ChEBI:CHEBI:136572"
FT /evidence="ECO:0000305|PubMed:11501996,
FT ECO:0007744|PDB:1JBW"
FT MOD_RES 185
FT /note="N6-carboxylysine"
FT /evidence="ECO:0000269|PubMed:11501996,
FT ECO:0007744|PDB:1JBV, ECO:0007744|PDB:1JBW"
FT MUTAGEN 151
FT /note="D->A: 220-fold decrease in catalytic efficiency with
FT mTHF as substrate, but only 4-fold decrease in catalytic
FT efficiency with 5,10-methylenetetrahydropteroyldiglutamate
FT as substrate."
FT /evidence="ECO:0000269|PubMed:18232714"
FT MUTAGEN 316
FT /note="H->A: Loss of activity."
FT /evidence="ECO:0000303|PubMed:11501996"
FT MUTAGEN 412
FT /note="S->A: Loss of activity."
FT /evidence="ECO:0000303|PubMed:11501996"
FT HELIX 3..10
FT /evidence="ECO:0007829|PDB:1JBW"
FT HELIX 23..31
FT /evidence="ECO:0007829|PDB:1JBW"
FT HELIX 35..37
FT /evidence="ECO:0007829|PDB:1JBW"
FT STRAND 41..45
FT /evidence="ECO:0007829|PDB:1JBW"
FT STRAND 47..49
FT /evidence="ECO:0007829|PDB:2GC5"
FT HELIX 50..63
FT /evidence="ECO:0007829|PDB:1JBW"
FT STRAND 68..71
FT /evidence="ECO:0007829|PDB:1JBW"
FT STRAND 76..78
FT /evidence="ECO:0007829|PDB:1FGS"
FT HELIX 79..82
FT /evidence="ECO:0007829|PDB:1JBW"
FT STRAND 83..85
FT /evidence="ECO:0007829|PDB:1JBW"
FT HELIX 92..112
FT /evidence="ECO:0007829|PDB:1JBW"
FT HELIX 120..134
FT /evidence="ECO:0007829|PDB:1JBW"
FT STRAND 138..143
FT /evidence="ECO:0007829|PDB:1JBW"
FT STRAND 145..148
FT /evidence="ECO:0007829|PDB:1JBW"
FT STRAND 159..163
FT /evidence="ECO:0007829|PDB:1JBW"
FT HELIX 168..170
FT /evidence="ECO:0007829|PDB:2GC5"
FT HELIX 171..174
FT /evidence="ECO:0007829|PDB:1JBW"
FT HELIX 178..185
FT /evidence="ECO:0007829|PDB:1JBW"
FT HELIX 186..188
FT /evidence="ECO:0007829|PDB:1JBW"
FT STRAND 195..197
FT /evidence="ECO:0007829|PDB:1JBW"
FT HELIX 202..215
FT /evidence="ECO:0007829|PDB:1JBW"
FT STRAND 219..221
FT /evidence="ECO:0007829|PDB:1JBW"
FT TURN 222..224
FT /evidence="ECO:0007829|PDB:1JBW"
FT STRAND 225..233
FT /evidence="ECO:0007829|PDB:1JBW"
FT STRAND 235..244
FT /evidence="ECO:0007829|PDB:1JBW"
FT STRAND 247..255
FT /evidence="ECO:0007829|PDB:1JBW"
FT HELIX 260..278
FT /evidence="ECO:0007829|PDB:1JBW"
FT HELIX 285..293
FT /evidence="ECO:0007829|PDB:1JBW"
FT STRAND 300..305
FT /evidence="ECO:0007829|PDB:1JBW"
FT TURN 306..309
FT /evidence="ECO:0007829|PDB:1JBW"
FT STRAND 310..313
FT /evidence="ECO:0007829|PDB:1JBW"
FT HELIX 318..331
FT /evidence="ECO:0007829|PDB:1JBW"
FT STRAND 337..341
FT /evidence="ECO:0007829|PDB:1JBW"
FT STRAND 343..345
FT /evidence="ECO:0007829|PDB:1JBW"
FT HELIX 348..358
FT /evidence="ECO:0007829|PDB:1JBW"
FT STRAND 360..364
FT /evidence="ECO:0007829|PDB:1JBW"
FT HELIX 391..401
FT /evidence="ECO:0007829|PDB:1JBW"
FT STRAND 407..412
FT /evidence="ECO:0007829|PDB:1JBW"
FT HELIX 413..423
FT /evidence="ECO:0007829|PDB:1JBW"
SQ SEQUENCE 428 AA; 46589 MW; E4B0B9ED62140181 CRC64;
MNYTETVAYI HSFPRLAKTG DHRRILTLLH ALGNPQQQGR YIHVTGTNGK GSAANAIAHV
LEASGLTVGL YTSPFIMRFN ERIMIDHEPI PDAALVNAVA FVRAALERLQ QQQADFNVTE
FEFITALGYW YFRQRQVDVA VIEVGIGGDT DSTNVITPVV SVLTEVALDH QKLLGHTITA
IAKHKAGIIK RGIPVVTGNL VPDAAAVVAA KVATTGSQWL RFDRDFSVPK AKLHGWGQRF
TYEDQDGRIS DLEVPLVGDY QQRNMAIAIQ TAKVYAKQTE WPLTPQNIRQ GLAASHWPAR
LEKISDTPLI VIDGAHNPDG INGLITALKQ LFSQPITVIA GILADKDYAA MADRLTAAFS
TVYLVPVPGT PRALPEAGYE ALHEGRLKDS WQEALAASLN DVPDQPIVIT GSLYLASAVR
QTLLGGKS
