COBU_SALTY
ID COBU_SALTY Reviewed; 181 AA.
AC Q05599;
DT 01-JUN-1994, integrated into UniProtKB/Swiss-Prot.
DT 01-OCT-1996, sequence version 2.
DT 03-AUG-2022, entry version 145.
DE RecName: Full=Bifunctional adenosylcobalamin biosynthesis protein CobU;
DE AltName: Full=Adenosylcobinamide kinase;
DE EC=2.7.1.156 {ECO:0000269|PubMed:10869342, ECO:0000269|PubMed:7559521};
DE AltName: Full=Adenosylcobinamide-phosphate guanylyltransferase;
DE EC=2.7.7.62 {ECO:0000269|PubMed:10869342, ECO:0000269|PubMed:7559521};
GN Name=cobU; OrderedLocusNames=STM2018;
OS Salmonella typhimurium (strain LT2 / SGSC1412 / ATCC 700720).
OC Bacteria; Proteobacteria; Gammaproteobacteria; Enterobacterales;
OC Enterobacteriaceae; Salmonella.
OX NCBI_TaxID=99287;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=LT2;
RX PubMed=8501034; DOI=10.1128/jb.175.11.3303-3316.1993;
RA Roth J.R., Lawrence J.G., Rubenfield M., Kieffer-Higgins S., Church G.M.;
RT "Characterization of the cobalamin (vitamin B12) biosynthetic genes of
RT Salmonella typhimurium.";
RL J. Bacteriol. 175:3303-3316(1993).
RN [2]
RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA].
RC STRAIN=LT2 / SGSC1412 / ATCC 700720;
RX PubMed=11677609; DOI=10.1038/35101614;
RA McClelland M., Sanderson K.E., Spieth J., Clifton S.W., Latreille P.,
RA Courtney L., Porwollik S., Ali J., Dante M., Du F., Hou S., Layman D.,
RA Leonard S., Nguyen C., Scott K., Holmes A., Grewal N., Mulvaney E.,
RA Ryan E., Sun H., Florea L., Miller W., Stoneking T., Nhan M., Waterston R.,
RA Wilson R.K.;
RT "Complete genome sequence of Salmonella enterica serovar Typhimurium LT2.";
RL Nature 413:852-856(2001).
RN [3]
RP PROTEIN SEQUENCE OF N-TERMINUS, FUNCTION, CATALYTIC ACTIVITY,
RP BIOPHYSICOCHEMICAL PROPERTIES, AND SUBSTRATE SPECIFICITY.
RC STRAIN=LT2;
RX PubMed=7559521; DOI=10.1074/jbc.270.40.23560;
RA O'Toole G.A., Escalante-Semerena J.C.;
RT "Purification and characterization of the bifunctional CobU enzyme of
RT Salmonella typhimurium LT2. Evidence for a CobU-GMP intermediate.";
RL J. Biol. Chem. 270:23560-23569(1995).
RN [4]
RP CATALYTIC ACTIVITY, AND MUTAGENESIS OF HIS-46 AND HIS-47.
RC STRAIN=LT2;
RX PubMed=10869342; DOI=10.1074/jbc.m000977200;
RA Thomas M.G., Thompson T.B., Rayment I., Escalante-Semerena J.C.;
RT "Analysis of the adenosylcobinamide kinase/adenosylcobinamide-phosphate
RT guanylyltransferase (CobU) enzyme of Salmonella typhimurium LT2.
RT Identification of residue His-46 as the site of guanylylation.";
RL J. Biol. Chem. 275:27576-27586(2000).
RN [5]
RP X-RAY CRYSTALLOGRAPHY (2.3 ANGSTROMS), AND SUBUNIT.
RC STRAIN=LT2;
RX PubMed=9601028; DOI=10.1021/bi973178f;
RA Thompson T.B., Thomas M.G., Escalante-Semerena J.C., Rayment I.;
RT "Three-dimensional structure of adenosylcobinamide
RT kinase/adenosylcobinamide phosphate guanylyltransferase from Salmonella
RT typhimurium determined to 2.3-A resolution.";
RL Biochemistry 37:7686-7695(1998).
RN [6]
RP X-RAY CRYSTALLOGRAPHY (2.2 ANGSTROMS) IN COMPLEX WITH GMP AND PHOSPHATE,
RP AND ACTIVE SITE.
RX PubMed=10529169; DOI=10.1021/bi990910x;
RA Thompson T.B., Thomas M.G., Escalante-Semerena J.C., Rayment I.;
RT "Three-dimensional structure of adenosylcobinamide
RT kinase/adenosylcobinamide phosphate guanylyltransferase (CobU) complexed
RT with GMP: evidence for a substrate-induced transferase active site.";
RL Biochemistry 38:12995-13005(1999).
CC -!- FUNCTION: Catalyzes ATP-dependent phosphorylation of adenosylcobinamide
CC and addition of GMP to adenosylcobinamide phosphate.
CC {ECO:0000269|PubMed:7559521}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=adenosylcob(III)inamide + GTP = adenosylcob(III)inamide
CC phosphate + GDP + H(+); Xref=Rhea:RHEA:15765, ChEBI:CHEBI:2480,
CC ChEBI:CHEBI:15378, ChEBI:CHEBI:37565, ChEBI:CHEBI:58189,
CC ChEBI:CHEBI:58502; EC=2.7.1.156;
CC Evidence={ECO:0000269|PubMed:10869342, ECO:0000269|PubMed:7559521};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=adenosylcob(III)inamide + ATP = adenosylcob(III)inamide
CC phosphate + ADP + H(+); Xref=Rhea:RHEA:15769, ChEBI:CHEBI:2480,
CC ChEBI:CHEBI:15378, ChEBI:CHEBI:30616, ChEBI:CHEBI:58502,
CC ChEBI:CHEBI:456216; EC=2.7.1.156;
CC Evidence={ECO:0000269|PubMed:10869342, ECO:0000269|PubMed:7559521};
CC -!- CATALYTIC ACTIVITY:
CC Reaction=adenosylcob(III)inamide phosphate + GTP + H(+) =
CC adenosylcob(III)inamide-GDP + diphosphate; Xref=Rhea:RHEA:22712,
CC ChEBI:CHEBI:15378, ChEBI:CHEBI:33019, ChEBI:CHEBI:37565,
CC ChEBI:CHEBI:58502, ChEBI:CHEBI:60487; EC=2.7.7.62;
CC Evidence={ECO:0000269|PubMed:10869342, ECO:0000269|PubMed:7559521};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=66 uM for adenosylcobinamide {ECO:0000269|PubMed:7559521};
CC KM=206 uM for ATP {ECO:0000269|PubMed:7559521};
CC KM=102 uM for adenosylcobinamide phosphate
CC {ECO:0000269|PubMed:7559521};
CC KM=21.3 uM for GTP {ECO:0000269|PubMed:7559521};
CC Note=kcat is 15.1 min(-1) for adenosylcobinamide kinase activity.
CC kcat is 5.6 min(-1) for guanylyltransferase activity.;
CC pH dependence:
CC Optimum pH is 8.8-9.0. {ECO:0000269|PubMed:7559521};
CC Temperature dependence:
CC Optimum temperature is 37 degrees Celsius.
CC {ECO:0000269|PubMed:7559521};
CC -!- PATHWAY: Cofactor biosynthesis; adenosylcobalamin biosynthesis;
CC adenosylcobalamin from cob(II)yrinate a,c-diamide: step 5/7.
CC -!- PATHWAY: Cofactor biosynthesis; adenosylcobalamin biosynthesis;
CC adenosylcobalamin from cob(II)yrinate a,c-diamide: step 6/7.
CC -!- SUBUNIT: Homotrimer. {ECO:0000269|PubMed:10529169,
CC ECO:0000269|PubMed:9601028}.
CC -!- SIMILARITY: Belongs to the CobU/CobP family. {ECO:0000305}.
CC -!- SEQUENCE CAUTION:
CC Sequence=AAA27269.1; Type=Erroneous initiation; Note=Truncated N-terminus.; Evidence={ECO:0000305};
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DR EMBL; L12006; AAA27269.1; ALT_INIT; Genomic_DNA.
DR EMBL; AE006468; AAL20922.1; -; Genomic_DNA.
DR RefSeq; NP_460963.1; NC_003197.2.
DR RefSeq; WP_000973145.1; NC_003197.2.
DR PDB; 1C9K; X-ray; 2.20 A; A/B/C=2-181.
DR PDB; 1CBU; X-ray; 2.30 A; A/B/C=2-181.
DR PDBsum; 1C9K; -.
DR PDBsum; 1CBU; -.
DR AlphaFoldDB; Q05599; -.
DR SMR; Q05599; -.
DR STRING; 99287.STM2018; -.
DR DrugBank; DB00115; Cyanocobalamin.
DR DrugBank; DB01972; Guanosine-5'-Monophosphate.
DR PaxDb; Q05599; -.
DR EnsemblBacteria; AAL20922; AAL20922; STM2018.
DR GeneID; 1253539; -.
DR KEGG; stm:STM2018; -.
DR PATRIC; fig|99287.12.peg.2140; -.
DR HOGENOM; CLU_094161_0_2_6; -.
DR OMA; DCLTVWL; -.
DR PhylomeDB; Q05599; -.
DR BioCyc; MetaCyc:MON-13215; -.
DR BioCyc; SENT99287:STM2018-MON; -.
DR UniPathway; UPA00148; UER00236.
DR UniPathway; UPA00148; UER00237.
DR EvolutionaryTrace; Q05599; -.
DR Proteomes; UP000001014; Chromosome.
DR GO; GO:0036429; F:adenosylcobinamide kinase (ATP-specific) activity; IEA:RHEA.
DR GO; GO:0036428; F:adenosylcobinamide kinase (GTP-specific) activity; IEA:RHEA.
DR GO; GO:0005524; F:ATP binding; IEA:UniProtKB-KW.
DR GO; GO:0008820; F:cobinamide phosphate guanylyltransferase activity; IEA:UniProtKB-EC.
DR GO; GO:0005525; F:GTP binding; IEA:UniProtKB-KW.
DR GO; GO:0009236; P:cobalamin biosynthetic process; IBA:GO_Central.
DR GO; GO:0016310; P:phosphorylation; IEA:UniProtKB-KW.
DR CDD; cd00544; CobU; 1.
DR Gene3D; 3.40.50.300; -; 1.
DR InterPro; IPR003203; Cobinamide_kinase/P_G-Trfase.
DR InterPro; IPR027417; P-loop_NTPase.
DR PANTHER; PTHR34848; PTHR34848; 1.
DR Pfam; PF02283; CobU; 1.
DR PIRSF; PIRSF006135; CobU; 1.
DR SUPFAM; SSF52540; SSF52540; 1.
PE 1: Evidence at protein level;
KW 3D-structure; ATP-binding; Cobalamin biosynthesis;
KW Direct protein sequencing; GTP-binding; Kinase; Nucleotide-binding;
KW Reference proteome; Transferase.
FT CHAIN 1..181
FT /note="Bifunctional adenosylcobalamin biosynthesis protein
FT CobU"
FT /id="PRO_0000089998"
FT ACT_SITE 47
FT /note="GMP-histidine intermediate"
FT /evidence="ECO:0000269|PubMed:10529169"
FT BINDING 7..14
FT /ligand="GTP"
FT /ligand_id="ChEBI:CHEBI:37565"
FT BINDING 31..33
FT /ligand="GTP"
FT /ligand_id="ChEBI:CHEBI:37565"
FT BINDING 48..51
FT /ligand="GTP"
FT /ligand_id="ChEBI:CHEBI:37565"
FT BINDING 59
FT /ligand="GTP"
FT /ligand_id="ChEBI:CHEBI:37565"
FT BINDING 81
FT /ligand="GTP"
FT /ligand_id="ChEBI:CHEBI:37565"
FT MUTAGEN 46
FT /note="H->A: 3-5 fold reduction in activity."
FT /evidence="ECO:0000269|PubMed:10869342"
FT MUTAGEN 46
FT /note="H->A: No activity; when associated with Ala-47."
FT /evidence="ECO:0000269|PubMed:10869342"
FT MUTAGEN 47
FT /note="H->A: Almost no residual activity."
FT /evidence="ECO:0000269|PubMed:10869342"
FT MUTAGEN 47
FT /note="H->A: No activity; when associated with Ala-46."
FT /evidence="ECO:0000269|PubMed:10869342"
FT MUTAGEN 47
FT /note="H->N: Very little residual activity."
FT /evidence="ECO:0000269|PubMed:10869342"
FT STRAND 3..7
FT /evidence="ECO:0007829|PDB:1C9K"
FT HELIX 13..21
FT /evidence="ECO:0007829|PDB:1C9K"
FT STRAND 25..31
FT /evidence="ECO:0007829|PDB:1C9K"
FT HELIX 38..41
FT /evidence="ECO:0007829|PDB:1C9K"
FT HELIX 43..49
FT /evidence="ECO:0007829|PDB:1C9K"
FT STRAND 55..58
FT /evidence="ECO:0007829|PDB:1C9K"
FT HELIX 64..66
FT /evidence="ECO:0007829|PDB:1C9K"
FT STRAND 77..81
FT /evidence="ECO:0007829|PDB:1C9K"
FT HELIX 83..93
FT /evidence="ECO:0007829|PDB:1C9K"
FT HELIX 100..102
FT /evidence="ECO:0007829|PDB:1CBU"
FT HELIX 105..125
FT /evidence="ECO:0007829|PDB:1C9K"
FT STRAND 128..133
FT /evidence="ECO:0007829|PDB:1C9K"
FT HELIX 146..165
FT /evidence="ECO:0007829|PDB:1C9K"
FT STRAND 167..173
FT /evidence="ECO:0007829|PDB:1C9K"
FT STRAND 176..179
FT /evidence="ECO:0007829|PDB:1C9K"
SQ SEQUENCE 181 AA; 19902 MW; F6F89892C6BEE091 CRC64;
MMILVTGGAR SGKSRHAEAL IGDAPQVLYI ATSQILDDEM AARIQHHKDG RPAHWRTAEC
WRHLDTLITA DLAPDDAILL ECITTMVTNL LFALGGENDP EQWDYAAMER AIDDEIQILI
AACQRCPAKV VLVTNEVGMG IVPENRLARH FRDIAGRVNQ RLAAAADEVW LVVSGIGVKI
K
