FOM3_STRWE
ID FOM3_STRWE Reviewed; 534 AA.
AC Q56184;
DT 13-APR-2016, integrated into UniProtKB/Swiss-Prot.
DT 01-NOV-1996, sequence version 1.
DT 03-AUG-2022, entry version 89.
DE RecName: Full=Cytidylyl-2-hydroxyethylphosphonate methyltransferase {ECO:0000305};
DE EC=2.1.1.308 {ECO:0000269|PubMed:28678474};
DE AltName: Full=MeCbl-dependent radical SAM C-methyltransferase Fom3 {ECO:0000303|PubMed:28678474};
GN Name=fom3 {ECO:0000303|PubMed:7500951};
OS Streptomyces wedmorensis.
OC Bacteria; Actinobacteria; Streptomycetales; Streptomycetaceae;
OC Streptomyces.
OX NCBI_TaxID=43759;
RN [1]
RP NUCLEOTIDE SEQUENCE [GENOMIC DNA].
RC STRAIN=144-91;
RX PubMed=7500951; DOI=10.1007/bf00290527;
RA Hidaka T., Goda M., Kuzuyama T., Takei N., Hidaka M., Seto H.;
RT "Cloning and nucleotide sequence of fosfomycin biosynthetic genes of
RT Streptomyces wedmorensis.";
RL Mol. Gen. Genet. 249:274-280(1995).
RN [2]
RP FUNCTION, PATHWAY, AND DISRUPTION PHENOTYPE.
RX PubMed=17220970; DOI=10.1039/b614678c;
RA Woodyer R.D., Li G., Zhao H., van der Donk W.A.;
RT "New insight into the mechanism of methyl transfer during the biosynthesis
RT of fosfomycin.";
RL Chem. Commun. (Camb.) 4:359-361(2007).
RN [3]
RP PRELIMINARY FUNCTION, COFACTOR, AND MUTAGENESIS OF CYS-282; CYS-286 AND
RP CYS-289.
RX PubMed=24370735; DOI=10.1016/j.abb.2013.12.004;
RA Allen K.D., Wang S.C.;
RT "Initial characterization of Fom3 from Streptomyces wedmorensis: The
RT methyltransferase in fosfomycin biosynthesis.";
RL Arch. Biochem. Biophys. 543:67-73(2014).
RN [4]
RP FUNCTION, CATALYTIC ACTIVITY, COFACTOR, BIOPHYSICOCHEMICAL PROPERTIES, AND
RP PATHWAY.
RC STRAIN=144-91;
RX PubMed=28678474; DOI=10.1021/acs.biochem.7b00472;
RA Sato S., Kudo F., Kim S.Y., Kuzuyama T., Eguchi T.;
RT "Methylcobalamin-dependent radical SAM C-methyltransferase Fom3 recognizes
RT cytidylyl-2-hydroxyethylphosphonate and catalyzes the nonstereoselective C-
RT methylation in fosfomycin biosynthesis.";
RL Biochemistry 56:3519-3522(2017).
RN [5]
RP PATHWAY.
RX PubMed=29345912; DOI=10.1021/acs.biochem.7b01281;
RA Blaszczyk A.J., Booker S.J.;
RT "A (re)discovery of the Fom3 substrate.";
RL Biochemistry 57:891-892(2018).
CC -!- FUNCTION: Involved in fosfomycin biosynthesis (PubMed:17220970,
CC PubMed:24370735, PubMed:28678474). Catalyzes the C-methylation of
CC cytidylyl-2-hydroxyethylphosphonate (HEP-CMP) to form cytidylyl-2-
CC hydroxypropylphosphonate (HPP-CMP). The C-methylation is not
CC stereoselective and the ratio of (S)- to (R)-HPP-CMP is almost equal in
CC vitro (PubMed:28678474). {ECO:0000269|PubMed:17220970,
CC ECO:0000269|PubMed:24370735, ECO:0000269|PubMed:28678474}.
CC -!- CATALYTIC ACTIVITY:
CC Reaction=AH2 + cytidine 5'-{[hydroxy(2-
CC hydroxyethyl)phosphonoyl]phosphate} + 2 S-adenosyl-L-methionine = 5'-
CC deoxyadenosine + A + cytidine 5'-{[hydroxy(2-
CC hydroxypropyl)phosphonoyl]phosphate} + 2 H(+) + L-methionine + S-
CC adenosyl-L-homocysteine; Xref=Rhea:RHEA:59072, ChEBI:CHEBI:13193,
CC ChEBI:CHEBI:15378, ChEBI:CHEBI:17319, ChEBI:CHEBI:17499,
CC ChEBI:CHEBI:57844, ChEBI:CHEBI:57856, ChEBI:CHEBI:59789,
CC ChEBI:CHEBI:142876, ChEBI:CHEBI:142877; EC=2.1.1.308;
CC Evidence={ECO:0000269|PubMed:28678474};
CC PhysiologicalDirection=left-to-right; Xref=Rhea:RHEA:59073;
CC Evidence={ECO:0000269|PubMed:28678474};
CC -!- COFACTOR:
CC Name=[4Fe-4S] cluster; Xref=ChEBI:CHEBI:49883;
CC Evidence={ECO:0000269|PubMed:24370735, ECO:0000269|PubMed:28678474};
CC Note=Binds 1 [4Fe-4S] cluster. The cluster is coordinated with 3
CC cysteines and an exchangeable S-adenosyl-L-methionine.
CC {ECO:0000269|PubMed:24370735};
CC -!- COFACTOR:
CC Name=methylcob(III)alamin; Xref=ChEBI:CHEBI:28115;
CC Evidence={ECO:0000269|PubMed:28678474};
CC -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC Kinetic parameters:
CC KM=113 uM for HEP-CMP {ECO:0000269|PubMed:28678474};
CC Note=kcat is 0.89 h(-1). {ECO:0000269|PubMed:28678474};
CC -!- PATHWAY: Antibiotic biosynthesis; fosfomycin biosynthesis.
CC {ECO:0000269|PubMed:17220970, ECO:0000269|PubMed:28678474,
CC ECO:0000305|PubMed:29345912}.
CC -!- DISRUPTION PHENOTYPE: Disruption mutant does not produce fosfomycin.
CC {ECO:0000269|PubMed:17220970}.
CC -!- MISCELLANEOUS: Was originally thought to catalyze the C-methylation of
CC 2-hydroxyethylphosphonate (2-HEP) to form 2-hydroxypropylphosphonate
CC (2-HPP). {ECO:0000303|PubMed:24370735}.
CC -!- SIMILARITY: Belongs to the radical SAM superfamily. {ECO:0000305}.
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DR EMBL; AB016934; BAA32490.1; -; Genomic_DNA.
DR PIR; S60205; S60205.
DR AlphaFoldDB; Q56184; -.
DR SMR; Q56184; -.
DR KEGG; ag:BAA32490; -.
DR BRENDA; 2.1.1.308; 6118.
DR UniPathway; UPA01071; -.
DR GO; GO:0051539; F:4 iron, 4 sulfur cluster binding; IEA:UniProtKB-KW.
DR GO; GO:0031419; F:cobalamin binding; IEA:UniProtKB-KW.
DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
DR GO; GO:0008168; F:methyltransferase activity; IEA:UniProtKB-KW.
DR GO; GO:0017000; P:antibiotic biosynthetic process; IEA:UniProtKB-KW.
DR GO; GO:0032259; P:methylation; IEA:UniProtKB-KW.
DR Gene3D; 3.80.30.20; -; 1.
DR InterPro; IPR006158; Cobalamin-bd.
DR InterPro; IPR036724; Cobalamin-bd_sf.
DR InterPro; IPR006638; Elp3/MiaB/NifB.
DR InterPro; IPR034529; Fom3-like.
DR InterPro; IPR020612; Methylthiotransferase_CS.
DR InterPro; IPR007197; rSAM.
DR InterPro; IPR023404; rSAM_horseshoe.
DR Pfam; PF02310; B12-binding; 1.
DR Pfam; PF04055; Radical_SAM; 1.
DR SFLD; SFLDF00403; phosphonoacetaldehyde_methylas; 1.
DR SFLD; SFLDS00029; Radical_SAM; 1.
DR SMART; SM00729; Elp3; 1.
DR SUPFAM; SSF52242; SSF52242; 1.
DR PROSITE; PS51332; B12_BINDING; 1.
DR PROSITE; PS51918; RADICAL_SAM; 1.
PE 1: Evidence at protein level;
KW 4Fe-4S; Antibiotic biosynthesis; Cobalamin; Cobalt; Iron; Iron-sulfur;
KW Metal-binding; Methyltransferase; S-adenosyl-L-methionine; Transferase.
FT CHAIN 1..534
FT /note="Cytidylyl-2-hydroxyethylphosphonate
FT methyltransferase"
FT /id="PRO_0000435951"
FT DOMAIN 38..195
FT /note="B12-binding"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU00666"
FT DOMAIN 268..496
FT /note="Radical SAM core"
FT /evidence="ECO:0000255|PROSITE-ProRule:PRU01266"
FT BINDING 282
FT /ligand="[4Fe-4S] cluster"
FT /ligand_id="ChEBI:CHEBI:49883"
FT /ligand_note="4Fe-4S-S-AdoMet"
FT /evidence="ECO:0000305|PubMed:24370735"
FT BINDING 286
FT /ligand="[4Fe-4S] cluster"
FT /ligand_id="ChEBI:CHEBI:49883"
FT /ligand_note="4Fe-4S-S-AdoMet"
FT /evidence="ECO:0000305|PubMed:24370735"
FT BINDING 289
FT /ligand="[4Fe-4S] cluster"
FT /ligand_id="ChEBI:CHEBI:49883"
FT /ligand_note="4Fe-4S-S-AdoMet"
FT /evidence="ECO:0000305|PubMed:24370735"
FT MUTAGEN 282
FT /note="C->A: Lacks the [4Fe-4S] cluster; when associated
FT with A-286 and A-289."
FT /evidence="ECO:0000269|PubMed:24370735"
FT MUTAGEN 286
FT /note="C->A: Lacks the [4Fe-4S] cluster; when associated
FT with A-282 and A-289."
FT /evidence="ECO:0000269|PubMed:24370735"
FT MUTAGEN 289
FT /note="C->A: Lacks the [4Fe-4S] cluster; when associated
FT with A-282 and A-286."
FT /evidence="ECO:0000269|PubMed:24370735"
SQ SEQUENCE 534 AA; 60374 MW; B057C98996461338 CRC64;
MTIGSLGSTE FALHGKPAIR WGDLPQRVGK PETRRYQKVL LLNPSATLFR HDLPRCTYPL
GLGYIAAVLE KYGYEVKILD VFAEGYYNAQ PVDGDDQFLR YGLSDDDIVK VMKEFGPDVV
GISSIFSNQA DNVHHLLKLA DLVTPEAVTA IGGAHARYFP KACLDDPNLD AVFLGEGEMT
FLLWMEHLNG NVSDDEVHGI AWRDRDGKVQ IKPELPLISS MRPEGPETGK SSPMLSMAGE
LDHIPFPAWH HYNMEKYFEI KAYQSPYTVG SRVGQLYTSR GCTAHCTFCT TTHFWGQKLR
RRSVQDVVDE VLRLRDEYGI DEFHIQDDNI TNDMDHAREL FRAFKEVGLP WATPQGTALW
RMDEELLDLM AESGAYQVTF AIESGVQRVL KELIKKPLNL ERTSHLIKYA RSLGMHVHGF
FIIGMPPMCG NAGESIEEMQ ASYDYAEEAG FSSASFFAAS PIVGSELLRE CIRQGFVDPE
ESLYRMTYKQ GIINVPGLWD GEEIAELAAK FNRDFNARRD RAYTPQKQWN ANQY
