MOBA_ECOLI
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 03-AUG-2022, entry version 169.
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; Proteobacteria; 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; P32173; -.
DR PRIDE; P32173; -.
DR EnsemblBacteria; AAC76855; AAC76855; b3857.
DR EnsemblBacteria; BAE77451; BAE77451; BAE77451.
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 PhylomeDB; P32173; -.
DR BioCyc; EcoCyc:EG11829-MON; -.
DR BioCyc; MetaCyc:EG11829-MON; -.
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 Gene3D; 3.90.550.10; -; 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 Pfam; PF12804; NTP_transf_3; 1.
DR SUPFAM; SSF53448; SSF53448; 1.
DR TIGRFAMs; TIGR02665; molyb_mobA; 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
