ID   MOBA_ECOLI              Reviewed;         194 AA.
AC   P32173; Q2M8F5; Q9LBV0;
DT   01-OCT-1993, integrated into UniProtKB/Swiss-Prot.
DT   01-OCT-1993, sequence version 1.
DT   24-JAN-2024, entry version 176.
DE   RecName: Full=Molybdenum cofactor guanylyltransferase;
DE            Short=MoCo guanylyltransferase;
DE            EC=2.7.7.77;
DE   AltName: Full=GTP:molybdopterin guanylyltransferase;
DE   AltName: Full=Mo-MPT guanylyltransferase;
DE   AltName: Full=Molybdopterin guanylyltransferase;
DE   AltName: Full=Molybdopterin-guanine dinucleotide biosynthesis protein A;
DE   AltName: Full=Molybdopterin-guanine dinucleotide synthase;
DE            Short=MGD synthase;
DE   AltName: Full=Protein FA;
GN   Name=mobA; Synonyms=chlB, mob, narB; OrderedLocusNames=b3857, JW3829;
OS   Escherichia coli (strain K12).
OC   Bacteria; Pseudomonadota; Gammaproteobacteria; Enterobacterales;
OC   Enterobacteriaceae; Escherichia.
OX   NCBI_TaxID=83333;
RN   [1]
RP   NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC   STRAIN=K12 / MG1655 / ATCC 47076;
RX   PubMed=8346018; DOI=10.1093/nar/21.15.3391;
RA   Plunkett G. III, Burland V., Daniels D.L., Blattner F.R.;
RT   "Analysis of the Escherichia coli genome. III. DNA sequence of the region
RT   from 87.2 to 89.2 minutes.";
RL   Nucleic Acids Res. 21:3391-3398(1993).
RN   [2]
RP   NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC   STRAIN=K12 / MG1655 / ATCC 47076;
RX   PubMed=9278503; DOI=10.1126/science.277.5331.1453;
RA   Blattner F.R., Plunkett G. III, Bloch C.A., Perna N.T., Burland V.,
RA   Riley M., Collado-Vides J., Glasner J.D., Rode C.K., Mayhew G.F.,
RA   Gregor J., Davis N.W., Kirkpatrick H.A., Goeden M.A., Rose D.J., Mau B.,
RA   Shao Y.;
RT   "The complete genome sequence of Escherichia coli K-12.";
RL   Science 277:1453-1462(1997).
RN   [3]
RP   NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC   STRAIN=K12 / W3110 / ATCC 27325 / DSM 5911;
RX   PubMed=16738553; DOI=10.1038/msb4100049;
RA   Hayashi K., Morooka N., Yamamoto Y., Fujita K., Isono K., Choi S.,
RA   Ohtsubo E., Baba T., Wanner B.L., Mori H., Horiuchi T.;
RT   "Highly accurate genome sequences of Escherichia coli K-12 strains MG1655
RT   and W3110.";
RL   Mol. Syst. Biol. 2:E1-E5(2006).
RN   [4]
RP   NUCLEOTIDE SEQUENCE [GENOMIC DNA], AND INDUCTION.
RC   STRAIN=K12;
RX   PubMed=7551035; DOI=10.1099/13500872-141-7-1663;
RA   Iobbi-Nivol C., Palmer T., Whitty P.W., McNairn E., Boxer D.H.;
RT   "The mob locus of Escherichia coli K12 required for molybdenum cofactor
RT   biosynthesis is expressed at very low levels.";
RL   Microbiology 141:1663-1671(1995).
RN   [5]
RP   PROTEIN SEQUENCE OF 1-5, FUNCTION IN MGD BIOSYNTHESIS, AND SUBUNIT.
RX   PubMed=8020507; DOI=10.1111/j.1432-1033.1994.tb18913.x;
RA   Palmer T., Vasishta A., Whitty P.W., Boxer D.H.;
RT   "Isolation of protein FA, a product of the mob locus required for
RT   molybdenum cofactor biosynthesis in Escherichia coli.";
RL   Eur. J. Biochem. 222:687-692(1994).
RN   [6]
RP   FUNCTION IN MGD BIOSYNTHESIS, AND DISRUPTION PHENOTYPE.
RC   STRAIN=RK4353;
RX   PubMed=1648082; DOI=10.1016/s0021-9258(18)98870-8;
RA   Johnson J.L., Indermaur L.W., Rajagopalan K.V.;
RT   "Molybdenum cofactor biosynthesis in Escherichia coli. Requirement of the
RT   chlB gene product for the formation of molybdopterin guanine
RT   dinucleotide.";
RL   J. Biol. Chem. 266:12140-12145(1991).
RN   [7]
RP   FUNCTION IN BIS(MGD) AND MGD BIOSYNTHESIS, CATALYTIC ACTIVITY, SUBSTRATE
RP   SPECIFICITY, AND COFACTOR.
RC   STRAIN=K12 / MC4100 / ATCC 35695 / DSM 6574;
RX   PubMed=10978348; DOI=10.1074/jbc.m007407200;
RA   Temple C.A., Rajagopalan K.V.;
RT   "Mechanism of assembly of the bis(molybdopterin guanine
RT   dinucleotide)molybdenum cofactor in Rhodobacter sphaeroides dimethyl
RT   sulfoxide reductase.";
RL   J. Biol. Chem. 275:40202-40210(2000).
RN   [8]
RP   INTERACTION WITH MOEA AND MOBB.
RC   STRAIN=K12 / MC4100 / ATCC 35695 / DSM 6574;
RX   PubMed=12372836; DOI=10.1074/jbc.m205806200;
RA   Magalon A., Frixon C., Pommier J., Giordano G., Blasco F.;
RT   "In vivo interactions between gene products involved in the final stages of
RT   molybdenum cofactor biosynthesis in Escherichia coli.";
RL   J. Biol. Chem. 277:48199-48204(2002).
RN   [9]
RP   FUNCTION, CATALYTIC ACTIVITY, KINETIC PARAMETERS, DOMAIN, AND MUTAGENESIS
RP   OF 12-LEU--GLY-14 AND 79-PRO--GLY-82.
RX   PubMed=21081498; DOI=10.1074/jbc.m110.155671;
RA   Neumann M., Seduk F., Iobbi-Nivol C., Leimkuhler S.;
RT   "Molybdopterin dinucleotide biosynthesis in Escherichia coli:
RT   identification of amino acid residues of molybdopterin dinucleotide
RT   transferases that determine specificity for binding of guanine or cytosine
RT   nucleotides.";
RL   J. Biol. Chem. 286:1400-1408(2011).
RN   [10]
RP   X-RAY CRYSTALLOGRAPHY (1.65 ANGSTROMS) OF APOENZYME AND IN COMPLEX WITH
RP   MN-GTP, COFACTOR, AND SUBUNIT.
RC   STRAIN=K12 / MC4100 / ATCC 35695 / DSM 6574;
RX   PubMed=10978347; DOI=10.1074/jbc.m007406200;
RA   Lake M.W., Temple C.A., Rajagopalan K.V., Schindelin H.;
RT   "The crystal structure of the Escherichia coli MobA protein provides
RT   insight into molybdopterin guanine dinucleotide biosynthesis.";
RL   J. Biol. Chem. 275:40211-40217(2000).
RN   [11]
RP   X-RAY CRYSTALLOGRAPHY (1.35 ANGSTROMS).
RC   STRAIN=K12;
RX   PubMed=11080634; DOI=10.1016/s0969-2126(00)00518-9;
RA   Stevenson C.E., Sargent F., Buchanan G., Palmer T., Lawson D.M.;
RT   "Crystal structure of the molybdenum cofactor biosynthesis protein MobA
RT   from Escherichia coli at near-atomic resolution.";
RL   Structure 8:1115-1125(2000).
RN   [12]
RP   X-RAY CRYSTALLOGRAPHY (1.65 ANGSTROMS) OF MUTANTS ALA-19; LEU-22; ASN-101;
RP   ASP-180 AND ASP-182, AND MUTAGENESIS OF GLY-15; ARG-19; GLY-22; LYS-25;
RP   GLY-78; GLY-82; ASP-101; ARG-156; ASN-180 AND ASN-182.
RC   STRAIN=K12;
RX   PubMed=12719427; DOI=10.1074/jbc.m302639200;
RA   Guse A., Stevenson C.E., Kuper J., Buchanan G., Schwarz G., Giordano G.,
RA   Magalon A., Mendel R.R., Lawson D.M., Palmer T.;
RT   "Biochemical and structural analysis of the molybdenum cofactor
RT   biosynthesis protein MobA.";
RL   J. Biol. Chem. 278:25302-25307(2003).
CC   -!- FUNCTION: Transfers a GMP moiety from GTP to Mo-molybdopterin (Mo-MPT)
CC       cofactor (Moco or molybdenum cofactor) to form Mo-molybdopterin guanine
CC       dinucleotide (Mo-MGD) cofactor. Is also involved in the biosynthesis of
CC       the bis-MGD form of the Moco cofactor (Mo-bisMGD) in which the metal is
CC       symmetrically ligated by the dithiolene groups of two MGD molecules. Is
CC       necessary and sufficient for the in vitro activation of the DMSOR
CC       molybdoenzyme that uses the Mo-bisMGD form of molybdenum cofactor,
CC       which implies formation and efficient insertion of the cofactor into
CC       the enzyme without the need of a chaperone. Is specific for GTP since
CC       other nucleotides such as ATP and GMP cannot be utilized.
CC       {ECO:0000269|PubMed:10978348, ECO:0000269|PubMed:1648082,
CC       ECO:0000269|PubMed:21081498, ECO:0000269|PubMed:8020507}.
CC   -!- CATALYTIC ACTIVITY:
CC       Reaction=GTP + H(+) + Mo-molybdopterin = diphosphate + Mo-molybdopterin
CC         guanine dinucleotide; Xref=Rhea:RHEA:34243, ChEBI:CHEBI:15378,
CC         ChEBI:CHEBI:33019, ChEBI:CHEBI:37565, ChEBI:CHEBI:71302,
CC         ChEBI:CHEBI:71310; EC=2.7.7.77;
CC         Evidence={ECO:0000269|PubMed:10978348, ECO:0000269|PubMed:21081498};
CC   -!- COFACTOR:
CC       Name=Mg(2+); Xref=ChEBI:CHEBI:18420;
CC         Evidence={ECO:0000269|PubMed:10978347, ECO:0000269|PubMed:10978348};
CC       Name=Mn(2+); Xref=ChEBI:CHEBI:29035;
CC         Evidence={ECO:0000269|PubMed:10978347, ECO:0000269|PubMed:10978348};
CC       Note=Both divalent cations appear to be equally efficient in an vitro
CC       reconstitution assay. {ECO:0000269|PubMed:10978347,
CC       ECO:0000269|PubMed:10978348};
CC   -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC       Kinetic parameters:
CC         KM=6.5 uM for GTP {ECO:0000269|PubMed:21081498};
CC   -!- SUBUNIT: Monomer. An equilibrium exists between a monomeric and
CC       oligomeric form of the enzyme, which could be an octamer; whether this
CC       oligomeric arrangement is of functional relevance is unclear. Interacts
CC       with MoeA and MobB in vivo. {ECO:0000269|PubMed:10978347,
CC       ECO:0000269|PubMed:12372836, ECO:0000269|PubMed:8020507}.
CC   -!- INTERACTION:
CC       P32173; P12281: moeA; NbExp=3; IntAct=EBI-1133881, EBI-554393;
CC   -!- SUBCELLULAR LOCATION: Cytoplasm.
CC   -!- INDUCTION: Is expressed at very low levels under both aerobic and
CC       anaerobic growth conditions. {ECO:0000269|PubMed:7551035}.
CC   -!- DOMAIN: The N-terminal domain determines nucleotide recognition and
CC       specific binding, while the C-terminal domain determines the specific
CC       binding to the target protein. When the N-terminal domain of MobA is
CC       fused to the C-terminal domain of MocA, comparable kinetic constants as
CC       wild-type MobA are obtained with GTP, and the activity with CTP is
CC       completely lost. Consistent results are obtained when the N-terminal
CC       domain of MocA is fused to the C-terminal domain of MobA: the kinetic
CC       constants with CTP are comparable with the ones found for wild-type
CC       MocA, although no activity with GTP is detected.
CC       {ECO:0000269|PubMed:21081498}.
CC   -!- DISRUPTION PHENOTYPE: Cells lacking this gene are chlorate-resistant,
CC       fail to synthesize MGD and accumulate elevated quantities of MPT.
CC       {ECO:0000269|PubMed:1648082}.
CC   -!- SIMILARITY: Belongs to the MobA family. {ECO:0000305}.
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DR   EMBL; L19201; AAB02992.1; -; Genomic_DNA.
DR   EMBL; U00096; AAC76855.1; -; Genomic_DNA.
DR   EMBL; AP009048; BAE77451.1; -; Genomic_DNA.
DR   PIR; S40803; S40803.
DR   RefSeq; NP_418294.1; NC_000913.3.
DR   RefSeq; WP_001052181.1; NZ_STEB01000017.1.
DR   PDB; 1E5K; X-ray; 1.35 A; A=1-194.
DR   PDB; 1FR9; X-ray; 1.65 A; A=1-194.
DR   PDB; 1FRW; X-ray; 1.75 A; A=1-194.
DR   PDB; 1H4C; X-ray; 1.65 A; A=1-194.
DR   PDB; 1H4D; X-ray; 1.74 A; A=1-194.
DR   PDB; 1H4E; X-ray; 1.65 A; A=1-194.
DR   PDB; 1HJJ; X-ray; 1.65 A; A=1-194.
DR   PDB; 1HJL; X-ray; 2.00 A; A=1-194.
DR   PDBsum; 1E5K; -.
DR   PDBsum; 1FR9; -.
DR   PDBsum; 1FRW; -.
DR   PDBsum; 1H4C; -.
DR   PDBsum; 1H4D; -.
DR   PDBsum; 1H4E; -.
DR   PDBsum; 1HJJ; -.
DR   PDBsum; 1HJL; -.
DR   AlphaFoldDB; P32173; -.
DR   SMR; P32173; -.
DR   BioGRID; 4260705; 3.
DR   DIP; DIP-10233N; -.
DR   IntAct; P32173; 11.
DR   STRING; 511145.b3857; -.
DR   DrugBank; DB04137; Guanosine-5'-Triphosphate.
DR   jPOST; P32173; -.
DR   PaxDb; 511145-b3857; -.
DR   EnsemblBacteria; AAC76855; AAC76855; b3857.
DR   GeneID; 948349; -.
DR   KEGG; ecj:JW3829; -.
DR   KEGG; eco:b3857; -.
DR   PATRIC; fig|1411691.4.peg.2858; -.
DR   EchoBASE; EB1776; -.
DR   eggNOG; COG0746; Bacteria.
DR   HOGENOM; CLU_055597_5_1_6; -.
DR   InParanoid; P32173; -.
DR   OMA; IDFWYAK; -.
DR   OrthoDB; 9788394at2; -.
DR   PhylomeDB; P32173; -.
DR   BioCyc; EcoCyc:EG11829-MONOMER; -.
DR   BioCyc; MetaCyc:EG11829-MONOMER; -.
DR   BRENDA; 2.7.7.77; 2026.
DR   SABIO-RK; P32173; -.
DR   EvolutionaryTrace; P32173; -.
DR   PRO; PR:P32173; -.
DR   Proteomes; UP000000318; Chromosome.
DR   Proteomes; UP000000625; Chromosome.
DR   GO; GO:0005737; C:cytoplasm; IEA:UniProtKB-SubCell.
DR   GO; GO:0005525; F:GTP binding; IEA:UniProtKB-UniRule.
DR   GO; GO:0000287; F:magnesium ion binding; IDA:EcoCyc.
DR   GO; GO:0061603; F:molybdenum cofactor guanylyltransferase activity; IDA:EcoCyc.
DR   GO; GO:0016779; F:nucleotidyltransferase activity; IBA:GO_Central.
DR   GO; GO:1902758; P:bis(molybdopterin guanine dinucleotide)molybdenum biosynthetic process; IMP:EcoCyc.
DR   CDD; cd02503; MobA; 1.
DR   HAMAP; MF_00316; MobA; 1.
DR   InterPro; IPR025877; MobA-like_NTP_Trfase.
DR   InterPro; IPR013482; Molybde_CF_guanTrfase.
DR   InterPro; IPR029044; Nucleotide-diphossugar_trans.
DR   NCBIfam; TIGR02665; molyb_mobA; 1.
DR   PANTHER; PTHR19136; MOLYBDENUM COFACTOR GUANYLYLTRANSFERASE; 1.
DR   PANTHER; PTHR19136:SF81; MOLYBDENUM COFACTOR GUANYLYLTRANSFERASE; 1.
DR   Pfam; PF12804; NTP_transf_3; 1.
DR   SUPFAM; SSF53448; Nucleotide-diphospho-sugar transferases; 1.
PE   1: Evidence at protein level;
KW   3D-structure; Cytoplasm; Direct protein sequencing; GTP-binding; Magnesium;
KW   Manganese; Metal-binding; Molybdenum cofactor biosynthesis;
KW   Nucleotide-binding; Reference proteome; Transferase.
FT   CHAIN           1..194
FT                   /note="Molybdenum cofactor guanylyltransferase"
FT                   /id="PRO_0000134887"
FT   BINDING         12..14
FT                   /ligand="GTP"
FT                   /ligand_id="ChEBI:CHEBI:37565"
FT   BINDING         25
FT                   /ligand="GTP"
FT                   /ligand_id="ChEBI:CHEBI:37565"
FT   BINDING         53
FT                   /ligand="GTP"
FT                   /ligand_id="ChEBI:CHEBI:37565"
FT   BINDING         71
FT                   /ligand="GTP"
FT                   /ligand_id="ChEBI:CHEBI:37565"
FT   BINDING         101
FT                   /ligand="GTP"
FT                   /ligand_id="ChEBI:CHEBI:37565"
FT   BINDING         101
FT                   /ligand="Mg(2+)"
FT                   /ligand_id="ChEBI:CHEBI:18420"
FT   MUTAGEN         12..14
FT                   /note="LAG->TAA: 7.5-fold decrease in affinity for GTP and
FT                   nearly no effect on catalytic activity. Displays a 3-fold
FT                   decrease in activity with GTP and gains a low activity with
FT                   CTP as substrate; when associated with 79-LLTS-82."
FT                   /evidence="ECO:0000269|PubMed:21081498"
FT   MUTAGEN         15
FT                   /note="G->L: Complete loss of catalytic activity. Still
FT                   capable of binding MPT and MGD and interacting with both
FT                   MoeA and MobB."
FT                   /evidence="ECO:0000269|PubMed:12719427"
FT   MUTAGEN         19
FT                   /note="R->A: Slight reduction in catalytic activity."
FT                   /evidence="ECO:0000269|PubMed:12719427"
FT   MUTAGEN         22
FT                   /note="G->L: Nearly no effect on catalytic activity."
FT                   /evidence="ECO:0000269|PubMed:12719427"
FT   MUTAGEN         25
FT                   /note="K->A: Marked reduction in catalytic activity. Still
FT                   capable of interacting with both MoeA and MobB."
FT                   /evidence="ECO:0000269|PubMed:12719427"
FT   MUTAGEN         78
FT                   /note="G->L: Nearly no effect on catalytic activity."
FT                   /evidence="ECO:0000269|PubMed:12719427"
FT   MUTAGEN         79..82
FT                   /note="PLAG->LLTS: 11-fold decrease in affinity for GTP and
FT                   nearly no effect on catalytic activity. Displays a 3-fold
FT                   decrease in activity with GTP and gains a low activity with
FT                   CTP as substrate; when associated with 12-TAA-14."
FT                   /evidence="ECO:0000269|PubMed:21081498"
FT   MUTAGEN         82
FT                   /note="G->L: Slight reduction in catalytic activity."
FT                   /evidence="ECO:0000269|PubMed:12719427"
FT   MUTAGEN         101
FT                   /note="D->A: Complete loss of catalytic activity."
FT                   /evidence="ECO:0000269|PubMed:12719427"
FT   MUTAGEN         101
FT                   /note="D->N: Marked reduction in catalytic activity. Still
FT                   capable of interacting with both MoeA and MobB."
FT                   /evidence="ECO:0000269|PubMed:12719427"
FT   MUTAGEN         156
FT                   /note="R->A: Nearly no effect on catalytic activity."
FT                   /evidence="ECO:0000269|PubMed:12719427"
FT   MUTAGEN         180
FT                   /note="N->D: Nearly no effect on catalytic activity."
FT                   /evidence="ECO:0000269|PubMed:12719427"
FT   MUTAGEN         182
FT                   /note="N->D: Nearly no effect on catalytic activity."
FT                   /evidence="ECO:0000269|PubMed:12719427"
FT   HELIX           3..5
FT                   /evidence="ECO:0007829|PDB:1FR9"
FT   STRAND          6..12
FT                   /evidence="ECO:0007829|PDB:1E5K"
FT   STRAND          18..20
FT                   /evidence="ECO:0007829|PDB:1E5K"
FT   HELIX           25..27
FT                   /evidence="ECO:0007829|PDB:1E5K"
FT   STRAND          28..30
FT                   /evidence="ECO:0007829|PDB:1E5K"
FT   HELIX           35..46
FT                   /evidence="ECO:0007829|PDB:1E5K"
FT   STRAND          50..53
FT                   /evidence="ECO:0007829|PDB:1E5K"
FT   STRAND          55..57
FT                   /evidence="ECO:0007829|PDB:1E5K"
FT   HELIX           58..62
FT                   /evidence="ECO:0007829|PDB:1E5K"
FT   STRAND          67..69
FT                   /evidence="ECO:0007829|PDB:1FR9"
FT   HELIX           79..89
FT                   /evidence="ECO:0007829|PDB:1E5K"
FT   STRAND          92..99
FT                   /evidence="ECO:0007829|PDB:1E5K"
FT   HELIX           109..115
FT                   /evidence="ECO:0007829|PDB:1E5K"
FT   TURN            116..119
FT                   /evidence="ECO:0007829|PDB:1FR9"
FT   STRAND          121..126
FT                   /evidence="ECO:0007829|PDB:1E5K"
FT   STRAND          131..139
FT                   /evidence="ECO:0007829|PDB:1E5K"
FT   HELIX           142..151
FT                   /evidence="ECO:0007829|PDB:1E5K"
FT   HELIX           157..163
FT                   /evidence="ECO:0007829|PDB:1E5K"
FT   STRAND          167..170
FT                   /evidence="ECO:0007829|PDB:1E5K"
FT   TURN            175..178
FT                   /evidence="ECO:0007829|PDB:1E5K"
FT   HELIX           184..188
FT                   /evidence="ECO:0007829|PDB:1E5K"
SQ   SEQUENCE   194 AA;  21643 MW;  B79B32DD7348DD48 CRC64;
     MNLMTTITGV VLAGGKARRM GGVDKGLLEL NGKPLWQHVA DALMTQLSHV VVNANRHQEI
     YQASGLKVIE DSLADYPGPL AGMLSVMQQE AGEWFLFCPC DTPYIPPDLA ARLNHQRKDA
     PVVWVHDGER DHPTIALVNR AIEPLLLEYL QAGERRVMVF MRLAGGHAVD FSDHKDAFVN
     VNTPEELARW QEKR
//