ID CYSG_ECOLI Reviewed; 457 AA. AC P0AEA8; P11098; P76685; Q2M734; DT 06-DEC-2005, integrated into UniProtKB/Swiss-Prot. DT 06-DEC-2005, sequence version 1. DT 27-MAR-2024, entry version 142. DE RecName: Full=Siroheme synthase {ECO:0000255|HAMAP-Rule:MF_01646, ECO:0000303|PubMed:8243665}; DE Includes: DE RecName: Full=Uroporphyrinogen-III C-methyltransferase {ECO:0000255|HAMAP-Rule:MF_01646}; DE Short=Urogen III methylase {ECO:0000255|HAMAP-Rule:MF_01646}; DE EC=2.1.1.107 {ECO:0000255|HAMAP-Rule:MF_01646, ECO:0000269|PubMed:2407234, ECO:0000269|PubMed:8573073, ECO:0000269|PubMed:9461500}; DE AltName: Full=SUMT {ECO:0000255|HAMAP-Rule:MF_01646}; DE AltName: Full=Uroporphyrinogen III methylase {ECO:0000255|HAMAP-Rule:MF_01646}; DE Short=UROM {ECO:0000255|HAMAP-Rule:MF_01646}; DE Includes: DE RecName: Full=Precorrin-2 dehydrogenase {ECO:0000255|HAMAP-Rule:MF_01646}; DE EC=1.3.1.76 {ECO:0000255|HAMAP-Rule:MF_01646, ECO:0000269|PubMed:8243665, ECO:0000269|PubMed:9461500}; DE Includes: DE RecName: Full=Sirohydrochlorin ferrochelatase {ECO:0000255|HAMAP-Rule:MF_01646}; DE EC=4.99.1.4 {ECO:0000255|HAMAP-Rule:MF_01646, ECO:0000269|PubMed:8243665, ECO:0000269|PubMed:9461500}; GN Name=cysG {ECO:0000255|HAMAP-Rule:MF_01646}; GN OrderedLocusNames=b3368, JW3331; OS Escherichia coli (strain K12). OC Bacteria; Pseudomonadota; Gammaproteobacteria; Enterobacterales; OC Enterobacteriaceae; Escherichia. OX NCBI_TaxID=83333; RN [1] RP NUCLEOTIDE SEQUENCE [GENOMIC DNA]. RC STRAIN=K12; RX PubMed=2543955; DOI=10.1093/nar/17.10.3865; RA Bell A.I., Gaston K.L., Cole J.A., Busby S.J.W.; RT "Cloning of binding sequences for the Escherichia coli transcription RT activators, FNR and CRP: location of bases involved in discrimination RT between FNR and CRP."; RL Nucleic Acids Res. 17:3865-3874(1989). RN [2] RP NUCLEOTIDE SEQUENCE [GENOMIC DNA]. RC STRAIN=K12; RX PubMed=2200672; DOI=10.1111/j.1432-1033.1990.tb19125.x; RA Peakman T., Crouzet J., Mayaux J.F., Busby S.J.W., Mohan S., Harborne N., RA Wootton J., Nicolson R., Cole J.A.; RT "Nucleotide sequence, organisation and structural analysis of the products RT of genes in the nirB-cysG region of the Escherichia coli K-12 chromosome."; RL Eur. J. Biochem. 191:315-323(1990). RN [3] 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 [4] 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 [5] RP PROTEIN SEQUENCE OF 1-18, AND FUNCTION. RX PubMed=2407558; DOI=10.1016/0014-5793(90)80640-5; RA Warren M.J., Stolowich N.J., Santander P.J., Roessner C.A., Sowa B.A., RA Scott A.I.; RT "Enzymatic synthesis of dihydrosirohydrochlorin (precorrin-2) and of a RT novel pyrrocorphin by uroporphyrinogen III methylase."; RL FEBS Lett. 261:76-80(1990). RN [6] RP FUNCTION AS A METHYLTRANSFERASE, CATALYTIC ACTIVITY, AND SUBUNIT. RX PubMed=2407234; DOI=10.1042/bj2650725; RA Warren M.J., Roessner C.A., Santander P.J., Scott A.I.; RT "The Escherichia coli cysG gene encodes S-adenosylmethionine-dependent RT uroporphyrinogen III methylase."; RL Biochem. J. 265:725-729(1990). RN [7] RP FUNCTION, AND CATALYTIC ACTIVITY. RX PubMed=8243665; DOI=10.1016/0014-5793(93)80438-z; RA Spencer J.B., Stolowich N.J., Roessner C.A., Scott A.I.; RT "The Escherichia coli cysG gene encodes the multifunctional protein, RT siroheme synthase."; RL FEBS Lett. 335:57-60(1993). RN [8] RP FUNCTION, AND CATALYTIC ACTIVITY. RX PubMed=8573073; DOI=10.1042/bj3130415; RA Woodcock S.C., Warren M.J.; RT "Evidence for a covalent intermediate in the S-adenosyl-L-methionine- RT dependent transmethylation reaction catalysed by sirohaem synthase."; RL Biochem. J. 313:415-421(1996). RN [9] RP FUNCTION, CATALYTIC ACTIVITY, AND MUTAGENESIS OF GLY-21; GLY-224; ASP-227; RP LYS-270; ARG-298; ASP-303 AND ARG-309. RX PubMed=9461500; DOI=10.1042/bj3300121; RA Woodcock S.C., Raux E., Levillayer F., Thermes C., Rambach A., Warren M.J.; RT "Effect of mutations in the transmethylase and dehydrogenase/chelatase RT domains of sirohaem synthase (CysG) on sirohaem and cobalamin RT biosynthesis."; RL Biochem. J. 330:121-129(1998). CC -!- FUNCTION: Multifunctional enzyme that catalyzes the SAM-dependent CC methylations of uroporphyrinogen III at position C-2 and C-7 to form CC precorrin-2 via precorrin-1. Then it catalyzes the NAD-dependent ring CC dehydrogenation of precorrin-2 to yield sirohydrochlorin. Finally, it CC catalyzes the ferrochelation of sirohydrochlorin to yield siroheme. CC {ECO:0000255|HAMAP-Rule:MF_01646, ECO:0000269|PubMed:2407234, CC ECO:0000269|PubMed:2407558, ECO:0000269|PubMed:8243665, CC ECO:0000269|PubMed:8573073, ECO:0000269|PubMed:9461500}. CC -!- CATALYTIC ACTIVITY: CC Reaction=2 S-adenosyl-L-methionine + uroporphyrinogen III = H(+) + CC precorrin-2 + 2 S-adenosyl-L-homocysteine; Xref=Rhea:RHEA:32459, CC ChEBI:CHEBI:15378, ChEBI:CHEBI:57308, ChEBI:CHEBI:57856, CC ChEBI:CHEBI:58827, ChEBI:CHEBI:59789; EC=2.1.1.107; CC Evidence={ECO:0000255|HAMAP-Rule:MF_01646, CC ECO:0000269|PubMed:2407234, ECO:0000269|PubMed:8573073, CC ECO:0000269|PubMed:9461500}; CC -!- CATALYTIC ACTIVITY: CC Reaction=NAD(+) + precorrin-2 = 2 H(+) + NADH + sirohydrochlorin; CC Xref=Rhea:RHEA:15613, ChEBI:CHEBI:15378, ChEBI:CHEBI:57540, CC ChEBI:CHEBI:57945, ChEBI:CHEBI:58351, ChEBI:CHEBI:58827; EC=1.3.1.76; CC Evidence={ECO:0000255|HAMAP-Rule:MF_01646, CC ECO:0000269|PubMed:8243665, ECO:0000269|PubMed:9461500}; CC -!- CATALYTIC ACTIVITY: CC Reaction=2 H(+) + siroheme = Fe(2+) + sirohydrochlorin; CC Xref=Rhea:RHEA:24360, ChEBI:CHEBI:15378, ChEBI:CHEBI:29033, CC ChEBI:CHEBI:58351, ChEBI:CHEBI:60052; EC=4.99.1.4; CC Evidence={ECO:0000255|HAMAP-Rule:MF_01646, CC ECO:0000269|PubMed:8243665, ECO:0000269|PubMed:9461500}; CC -!- PATHWAY: Cofactor biosynthesis; adenosylcobalamin biosynthesis; CC precorrin-2 from uroporphyrinogen III: step 1/1. {ECO:0000255|HAMAP- CC Rule:MF_01646, ECO:0000305|PubMed:2407234}. CC -!- PATHWAY: Cofactor biosynthesis; adenosylcobalamin biosynthesis; CC sirohydrochlorin from precorrin-2: step 1/1. {ECO:0000255|HAMAP- CC Rule:MF_01646, ECO:0000305|PubMed:8243665}. CC -!- PATHWAY: Porphyrin-containing compound metabolism; siroheme CC biosynthesis; precorrin-2 from uroporphyrinogen III: step 1/1. CC {ECO:0000255|HAMAP-Rule:MF_01646, ECO:0000305|PubMed:2407234}. CC -!- PATHWAY: Porphyrin-containing compound metabolism; siroheme CC biosynthesis; siroheme from sirohydrochlorin: step 1/1. CC {ECO:0000255|HAMAP-Rule:MF_01646, ECO:0000305|PubMed:8243665}. CC -!- PATHWAY: Porphyrin-containing compound metabolism; siroheme CC biosynthesis; sirohydrochlorin from precorrin-2: step 1/1. CC {ECO:0000255|HAMAP-Rule:MF_01646, ECO:0000305|PubMed:8243665}. CC -!- SUBUNIT: Monomer. {ECO:0000269|PubMed:2407234}. CC -!- SIMILARITY: In the N-terminal section; belongs to the precorrin-2 CC dehydrogenase / sirohydrochlorin ferrochelatase family. CC {ECO:0000255|HAMAP-Rule:MF_01646}. CC -!- SIMILARITY: In the C-terminal section; belongs to the precorrin CC methyltransferase family. {ECO:0000255|HAMAP-Rule:MF_01646}. CC --------------------------------------------------------------------------- CC Copyrighted by the UniProt Consortium, see https://www.uniprot.org/terms CC Distributed under the Creative Commons Attribution (CC BY 4.0) License CC --------------------------------------------------------------------------- DR EMBL; X14202; CAA32419.1; -; Genomic_DNA. DR EMBL; U18997; AAA58165.1; -; Genomic_DNA. DR EMBL; U00096; AAC76393.1; -; Genomic_DNA. DR EMBL; AP009048; BAE77922.1; -; Genomic_DNA. DR PIR; C65131; C65131. DR RefSeq; NP_417827.1; NC_000913.3. DR RefSeq; WP_000349855.1; NZ_STEB01000004.1. DR AlphaFoldDB; P0AEA8; -. DR SMR; P0AEA8; -. DR BioGRID; 4262478; 6. DR IntAct; P0AEA8; 1. DR STRING; 511145.b3368; -. DR jPOST; P0AEA8; -. DR PaxDb; 511145-b3368; -. DR EnsemblBacteria; AAC76393; AAC76393; b3368. DR GeneID; 75173526; -. DR GeneID; 947880; -. DR KEGG; ecj:JW3331; -. DR KEGG; eco:b3368; -. DR PATRIC; fig|1411691.4.peg.3361; -. DR EchoBASE; EB0185; -. DR eggNOG; COG0007; Bacteria. DR eggNOG; COG1648; Bacteria. DR HOGENOM; CLU_011276_2_0_6; -. DR InParanoid; P0AEA8; -. DR OMA; IPYGRFM; -. DR OrthoDB; 9815856at2; -. DR PhylomeDB; P0AEA8; -. DR BioCyc; EcoCyc:SIROHEMESYN-MONOMER; -. DR BioCyc; MetaCyc:SIROHEMESYN-MONOMER; -. DR UniPathway; UPA00148; UER00211. DR UniPathway; UPA00148; UER00222. DR UniPathway; UPA00262; UER00211. DR UniPathway; UPA00262; UER00222. DR UniPathway; UPA00262; UER00376. DR PRO; PR:P0AEA8; -. DR Proteomes; UP000000318; Chromosome. DR Proteomes; UP000000625; Chromosome. DR GO; GO:0051287; F:NAD binding; IEA:InterPro. DR GO; GO:0043115; F:precorrin-2 dehydrogenase activity; IEA:UniProtKB-UniRule. DR GO; GO:0042803; F:protein homodimerization activity; IPI:EcoCyc. DR GO; GO:0051266; F:sirohydrochlorin ferrochelatase activity; IEA:UniProtKB-EC. DR GO; GO:0004851; F:uroporphyrin-III C-methyltransferase activity; IDA:EcoCyc. DR GO; GO:0009236; P:cobalamin biosynthetic process; IEA:UniProtKB-UniRule. DR GO; GO:0032259; P:methylation; IEA:UniProtKB-KW. DR GO; GO:0006970; P:response to osmotic stress; IMP:EcoCyc. DR GO; GO:0019354; P:siroheme biosynthetic process; IBA:GO_Central. DR CDD; cd11642; SUMT; 1. DR Gene3D; 3.40.50.720; NAD(P)-binding Rossmann-like Domain; 1. DR Gene3D; 1.10.8.210; Sirohaem synthase, dimerisation domain; 1. DR HAMAP; MF_01646; Siroheme_synth; 1. DR InterPro; IPR000878; 4pyrrol_Mease. DR InterPro; IPR035996; 4pyrrol_Methylase_sf. DR InterPro; IPR014777; 4pyrrole_Mease_sub1. DR InterPro; IPR014776; 4pyrrole_Mease_sub2. DR InterPro; IPR006366; CobA/CysG_C. DR InterPro; IPR036291; NAD(P)-bd_dom_sf. DR InterPro; IPR037115; Sirohaem_synt_dimer_dom_sf. DR InterPro; IPR012409; Sirohaem_synth. DR InterPro; IPR028281; Sirohaem_synthase_central. DR InterPro; IPR019478; Sirohaem_synthase_dimer_dom. DR InterPro; IPR006367; Sirohaem_synthase_N. DR InterPro; IPR003043; Uropor_MeTrfase_CS. DR NCBIfam; TIGR01469; cobA_cysG_Cterm; 1. DR NCBIfam; TIGR01470; cysG_Nterm; 1. DR PANTHER; PTHR45790:SF1; SIROHEME SYNTHASE; 1. DR PANTHER; PTHR45790; SIROHEME SYNTHASE-RELATED; 1. DR Pfam; PF10414; CysG_dimeriser; 1. DR Pfam; PF13241; NAD_binding_7; 1. DR Pfam; PF14824; Sirohm_synth_M; 1. DR Pfam; PF00590; TP_methylase; 1. DR PIRSF; PIRSF036426; Sirohaem_synth; 1. DR SUPFAM; SSF51735; NAD(P)-binding Rossmann-fold domains; 1. DR SUPFAM; SSF75615; Siroheme synthase middle domains-like; 1. DR SUPFAM; SSF53790; Tetrapyrrole methylase; 1. DR PROSITE; PS00839; SUMT_1; 1. DR PROSITE; PS00840; SUMT_2; 1. PE 1: Evidence at protein level; KW Cobalamin biosynthesis; Direct protein sequencing; Lyase; KW Methyltransferase; Multifunctional enzyme; NAD; Oxidoreductase; KW Phosphoprotein; Porphyrin biosynthesis; Reference proteome; KW S-adenosyl-L-methionine; Transferase. FT CHAIN 1..457 FT /note="Siroheme synthase" FT /id="PRO_0000150378" FT REGION 4..204 FT /note="Precorrin-2 dehydrogenase /sirohydrochlorin FT ferrochelatase" FT /evidence="ECO:0000250" FT REGION 216..448 FT /note="Uroporphyrinogen-III C-methyltransferase" FT /evidence="ECO:0000255|HAMAP-Rule:MF_01646" FT ACT_SITE 248 FT /note="Proton acceptor" FT /evidence="ECO:0000255|HAMAP-Rule:MF_01646" FT ACT_SITE 270 FT /note="Proton donor" FT /evidence="ECO:0000255|HAMAP-Rule:MF_01646" FT BINDING 22..23 FT /ligand="NAD(+)" FT /ligand_id="ChEBI:CHEBI:57540" FT /evidence="ECO:0000255|HAMAP-Rule:MF_01646" FT BINDING 43..44 FT /ligand="NAD(+)" FT /ligand_id="ChEBI:CHEBI:57540" FT /evidence="ECO:0000255|HAMAP-Rule:MF_01646" FT BINDING 225 FT /ligand="S-adenosyl-L-methionine" FT /ligand_id="ChEBI:CHEBI:59789" FT /evidence="ECO:0000255|HAMAP-Rule:MF_01646" FT BINDING 301..303 FT /ligand="S-adenosyl-L-methionine" FT /ligand_id="ChEBI:CHEBI:59789" FT /evidence="ECO:0000255|HAMAP-Rule:MF_01646" FT BINDING 306 FT /ligand="S-adenosyl-L-methionine" FT /ligand_id="ChEBI:CHEBI:59789" FT /evidence="ECO:0000255|HAMAP-Rule:MF_01646" FT BINDING 331..332 FT /ligand="S-adenosyl-L-methionine" FT /ligand_id="ChEBI:CHEBI:59789" FT /evidence="ECO:0000255|HAMAP-Rule:MF_01646" FT BINDING 382 FT /ligand="S-adenosyl-L-methionine" FT /ligand_id="ChEBI:CHEBI:59789" FT /evidence="ECO:0000255|HAMAP-Rule:MF_01646" FT BINDING 411 FT /ligand="S-adenosyl-L-methionine" FT /ligand_id="ChEBI:CHEBI:59789" FT /evidence="ECO:0000255|HAMAP-Rule:MF_01646" FT MOD_RES 128 FT /note="Phosphoserine" FT /evidence="ECO:0000255|HAMAP-Rule:MF_01646" FT MUTAGEN 21 FT /note="G->D: It has methyltransferase and ferrochelatase FT activities but a greatly reduced dehydrogenase activity. It FT is able to bind AdoMet." FT /evidence="ECO:0000269|PubMed:9461500" FT MUTAGEN 224 FT /note="G->A: It abolishes methyltransferase activity, but FT has dehydrogenase and ferrochelatase activities. It is FT unable to bind AdoMet." FT /evidence="ECO:0000269|PubMed:9461500" FT MUTAGEN 227 FT /note="D->A: It has all activities of CysG." FT /evidence="ECO:0000269|PubMed:9461500" FT MUTAGEN 248 FT /note="D->A: It abolishes methyltransferase activity, but FT has dehydrogenase and ferrochelatase activities. It is able FT to bind AdoMet." FT MUTAGEN 270 FT /note="K->I: It has all activities of CysG. It is able to FT bind AdoMet." FT /evidence="ECO:0000269|PubMed:9461500" FT MUTAGEN 298 FT /note="R->L: It abolishes methyltransferase activity, but FT has dehydrogenase and ferrochelatase activities. It is FT unable to bind AdoMet." FT /evidence="ECO:0000269|PubMed:9461500" FT MUTAGEN 303 FT /note="D->A: It has all activities of CysG. It is able to FT bind AdoMet." FT /evidence="ECO:0000269|PubMed:9461500" FT MUTAGEN 309 FT /note="R->L: It abolishes methyltransferase activity, but FT has dehydrogenase and ferrochelatase activities. It is able FT to bind AdoMet." FT /evidence="ECO:0000269|PubMed:9461500" FT CONFLICT 26 FT /note="R -> P (in Ref. 1; CAA32419 and 2; AAA58165)" FT /evidence="ECO:0000305" FT CONFLICT 90..91 FT /note="SE -> RQ (in Ref. 1; CAA32419 and 2; AAA58165)" FT /evidence="ECO:0000305" SQ SEQUENCE 457 AA; 49951 MW; DDDE0DC90C5CF2F4 CRC64; MDHLPIFCQL RDRDCLIVGG GDVAERKARL LLDAGARLTV NALAFIPQFT AWADAGMLTL VEGPFDESLL DTCWLAIAAT DDDALNQRVS EAAEARRIFC NVVDAPKAAS FIMPSIIDRS PLMVAVSSGG TSPVLARLLR EKLESLLPLH LGQVAKYAGQ LRGRVKQQFA TMGERRRFWE KLFVNDRLAQ SLANNDQKAI TETTEQLINE PLDHRGEVVL VGAGPGDAGL LTLKGLQQIQ QADVVVYDRL VSDDIMNLVR RDADRVFVGK RAGYHCVPQE EINQILLREA QKGKRVVRLK GGDPFIFGRG GEELETLCNA GIPFSVVPGI TAASGCSAYS GIPLTHRDYA QSVRLITGHL KTGGELDWEN LAAEKQTLVF YMGLNQAATI QQKLIEHGMP GEMPVAIVEN GTAVTQRVID GTLTQLGELA QQMNSPSLII IGRVVGLRDK LNWFSNH //