ID   MGM_EUBBA               Reviewed;         614 AA.
AC   Q59268; Q0QLE7;
DT   28-NOV-2002, integrated into UniProtKB/Swiss-Prot.
DT   01-NOV-1996, sequence version 1.
DT   27-MAR-2024, entry version 93.
DE   RecName: Full=2-methyleneglutarate mutase;
DE            EC=5.4.99.4;
DE   AltName: Full=Alpha-methyleneglutarate mutase;
GN   Name=mgm;
OS   Eubacterium barkeri (Clostridium barkeri).
OC   Bacteria; Bacillota; Clostridia; Eubacteriales; Eubacteriaceae;
OC   Eubacterium.
OX   NCBI_TaxID=1528;
RN   [1]
RP   NUCLEOTIDE SEQUENCE [GENOMIC DNA], PROTEIN SEQUENCE OF 1-21; 68-78; 79-103;
RP   122-126; 141-145; 395-409; 494-507; 547-552 AND 564-576, CATALYTIC
RP   ACTIVITY, AND COFACTOR.
RC   STRAIN=ATCC 25849 / DSM 1223 / JCM 1389 / NCIMB 10623 / VKM B-1775 /
RC   VPI 5359;
RX   PubMed=8168499; DOI=10.1111/j.1432-1033.1994.tb18718.x;
RA   Beatrix B., Zelder O., Linder D., Buckel W.;
RT   "Cloning, sequencing and expression of the gene encoding the coenzyme B12-
RT   dependent 2-methyleneglutarate mutase from Clostridium barkeri in
RT   Escherichia coli.";
RL   Eur. J. Biochem. 221:101-109(1994).
RN   [2]
RP   NUCLEOTIDE SEQUENCE [GENOMIC DNA], AND PATHWAY.
RC   STRAIN=ATCC 25849 / DSM 1223 / JCM 1389 / NCIMB 10623 / VKM B-1775 /
RC   VPI 5359;
RX   PubMed=16894175; DOI=10.1073/pnas.0601635103;
RA   Alhapel A., Darley D.J., Wagener N., Eckel E., Elsner N., Pierik A.J.;
RT   "Molecular and functional analysis of nicotinate catabolism in Eubacterium
RT   barkeri.";
RL   Proc. Natl. Acad. Sci. U.S.A. 103:12341-12346(2006).
RN   [3]
RP   CATALYTIC ACTIVITY, COFACTOR, ACTIVITY REGULATION, BIOPHYSICOCHEMICAL
RP   PROPERTIES, AND INDUCTION.
RX   PubMed=5574401; DOI=10.1016/s0021-9258(18)62235-5;
RA   Kung H.F., Stadtman T.C.;
RT   "Nicotinic acid metabolism. VI. Purification and properties of alpha-
RT   methyleneglutarate mutase (B 12-dependent) and methylitaconate isomerase.";
RL   J. Biol. Chem. 246:3378-3388(1971).
RN   [4]
RP   CATALYTIC ACTIVITY, AND SUBUNIT.
RX   PubMed=2776761; DOI=10.1111/j.1432-1033.1989.tb14995.x;
RA   Michel C., Hartrampf G., Buckel W.;
RT   "Assay and purification of the adenosylcobalamin-dependent 2-
RT   methyleneglutarate mutase from Clostridium barkeri.";
RL   Eur. J. Biochem. 184:103-107(1989).
RN   [5]
RP   COFACTOR.
RX   PubMed=1315277; DOI=10.1111/j.1432-1033.1992.tb16841.x;
RA   Michel C., Albracht S.P., Buckel W.;
RT   "Adenosylcobalamin and cob(II)alamin as prosthetic groups of 2-
RT   methyleneglutarate mutase from Clostridium barkeri.";
RL   Eur. J. Biochem. 205:767-773(1992).
RN   [6]
RP   COFACTOR.
RX   PubMed=8382495; DOI=10.1515/bchm3.1993.374.1-6.85;
RA   Zelder O., Buckel W.;
RT   "On the role of two different cobalt(II) species in coenzyme B12-dependent
RT   2-methyleneglutarate mutase from Clostridium barkeri.";
RL   Biol. Chem. Hoppe-Seyler 374:85-90(1993).
RN   [7]
RP   CATALYTIC ACTIVITY, COFACTOR, ACTIVITY REGULATION, AND MUTAGENESIS OF
RP   HIS-464; ASP-483 AND HIS-485.
RX   PubMed=16060663; DOI=10.1021/bi050049n;
RA   Pierik A.J., Ciceri D., Lopez R.F., Kroll F., Broker G., Beatrix B.,
RA   Buckel W., Golding B.T.;
RT   "Searching for intermediates in the carbon skeleton rearrangement of 2-
RT   methyleneglutarate to (R)-3-methylitaconate catalyzed by coenzyme B12-
RT   dependent 2-methyleneglutarate mutase from Eubacterium barkeri.";
RL   Biochemistry 44:10541-10551(2005).
CC   -!- FUNCTION: Involved in the fermentation of nicotinate to ammonia,
CC       propionate, acetate and carbon dioxide.
CC   -!- CATALYTIC ACTIVITY:
CC       Reaction=2-methyleneglutarate = 2-methylene-3-methylsuccinate;
CC         Xref=Rhea:RHEA:13793, ChEBI:CHEBI:57637, ChEBI:CHEBI:58056;
CC         EC=5.4.99.4; Evidence={ECO:0000269|PubMed:16060663,
CC         ECO:0000269|PubMed:2776761, ECO:0000269|PubMed:5574401,
CC         ECO:0000269|PubMed:8168499};
CC   -!- COFACTOR:
CC       Name=adenosylcob(III)alamin; Xref=ChEBI:CHEBI:18408;
CC         Evidence={ECO:0000269|PubMed:1315277, ECO:0000269|PubMed:16060663,
CC         ECO:0000269|PubMed:5574401, ECO:0000269|PubMed:8168499,
CC         ECO:0000269|PubMed:8382495};
CC       Name=cob(II)alamin; Xref=ChEBI:CHEBI:16304;
CC         Evidence={ECO:0000269|PubMed:1315277, ECO:0000269|PubMed:16060663,
CC         ECO:0000269|PubMed:5574401, ECO:0000269|PubMed:8168499,
CC         ECO:0000269|PubMed:8382495};
CC       Note=Contains a mixture of adenosylcobalamin and oxygen-stable
CC       cob(II)alamin. {ECO:0000269|PubMed:1315277,
CC       ECO:0000269|PubMed:16060663, ECO:0000269|PubMed:5574401,
CC       ECO:0000269|PubMed:8168499, ECO:0000269|PubMed:8382495};
CC   -!- ACTIVITY REGULATION: Inhibited by intrinsic factor and iodoacetic acid.
CC       Competitively inhibited by itaconate, mesaconate, succinate, 1-methyl-
CC       1,2-trans-clycopropane, dicarboxylate and L-malate. Non-competitively
CC       inhibited by glutaconate and 1-methyl-1,2-cis-
CC       cyclopropanedicarboxylate. Not inhibited by acrylate or by the
CC       chelating agents alpha,alpha-dipyridyl or EDTA. Not activated by
CC       Fe(2+), Mg(2+), Mn(2+) or Ca(2+). Unaffected by K(+), Na(+), NH4(+),
CC       Rb(+) or Li(+). {ECO:0000269|PubMed:16060663,
CC       ECO:0000269|PubMed:5574401}.
CC   -!- BIOPHYSICOCHEMICAL PROPERTIES:
CC       pH dependence:
CC         Activity decreases rapidly below pH 7.5 in phosphate buffer, and
CC         below pH 8.0 in Tris buffer. {ECO:0000269|PubMed:5574401};
CC   -!- PATHWAY: Cofactor degradation; nicotinate degradation; propanoate and
CC       pyruvate from 6-hydroxynicotinate: step 5/8.
CC       {ECO:0000269|PubMed:16894175}.
CC   -!- SUBUNIT: Homotetramer. {ECO:0000269|PubMed:2776761}.
CC   -!- INDUCTION: By nicotinate. {ECO:0000269|PubMed:5574401}.
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DR   EMBL; X77484; CAA54625.1; -; Genomic_DNA.
DR   EMBL; DQ310789; ABC88403.1; -; Genomic_DNA.
DR   PIR; S43237; S43237.
DR   AlphaFoldDB; Q59268; -.
DR   SMR; Q59268; -.
DR   STRING; 1528.SAMN04488579_1129; -.
DR   KEGG; ag:CAA54625; -.
DR   OrthoDB; 1672287at2; -.
DR   BioCyc; MetaCyc:MONOMER-11712; -.
DR   UniPathway; UPA01010; UER01016.
DR   GO; GO:0047548; F:2-methyleneglutarate mutase activity; IDA:UniProtKB.
DR   GO; GO:0031419; F:cobalamin binding; IEA:UniProtKB-KW.
DR   GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW.
DR   GO; GO:1901848; P:nicotinate catabolic process; IDA:UniProtKB.
DR   CDD; cd02071; MM_CoA_mut_B12_BD; 1.
DR   Gene3D; 3.40.50.280; Cobalamin-binding domain; 1.
DR   Gene3D; 3.20.20.240; Methylmalonyl-CoA mutase; 1.
DR   InterPro; IPR006159; Acid_CoA_mut_C.
DR   InterPro; IPR016176; Cbl-dep_enz_cat.
DR   InterPro; IPR006158; Cobalamin-bd.
DR   InterPro; IPR036724; Cobalamin-bd_sf.
DR   NCBIfam; TIGR00640; acid_CoA_mut_C; 1.
DR   PANTHER; PTHR48101:SF1; METHYLMALONYL-COA MUTASE, LARGE SUBUNIT; 1.
DR   PANTHER; PTHR48101; METHYLMALONYL-COA MUTASE, MITOCHONDRIAL-RELATED; 1.
DR   Pfam; PF02310; B12-binding; 1.
DR   SUPFAM; SSF52242; Cobalamin (vitamin B12)-binding domain; 1.
DR   SUPFAM; SSF51703; Cobalamin (vitamin B12)-dependent enzymes; 1.
DR   PROSITE; PS51332; B12_BINDING; 1.
PE   1: Evidence at protein level;
KW   Cobalamin; Cobalt; Direct protein sequencing; Isomerase; Metal-binding.
FT   CHAIN           1..614
FT                   /note="2-methyleneglutarate mutase"
FT                   /id="PRO_0000096464"
FT   DOMAIN          472..614
FT                   /note="B12-binding"
FT                   /evidence="ECO:0000255|PROSITE-ProRule:PRU00666"
FT   BINDING         485
FT                   /ligand="adenosylcob(III)alamin"
FT                   /ligand_id="ChEBI:CHEBI:18408"
FT                   /ligand_part="Co"
FT                   /ligand_part_id="ChEBI:CHEBI:27638"
FT                   /note="axial binding residue"
FT                   /evidence="ECO:0000250"
FT   MUTAGEN         464
FT                   /note="H->Q: No effect on activity."
FT                   /evidence="ECO:0000269|PubMed:16060663"
FT   MUTAGEN         483
FT                   /note="D->N: Activity reduced 2000-fold."
FT                   /evidence="ECO:0000269|PubMed:16060663"
FT   MUTAGEN         485
FT                   /note="H->Q: Loss of activity."
FT                   /evidence="ECO:0000269|PubMed:16060663"
FT   CONFLICT        97
FT                   /note="K -> W (in Ref. 1; AA sequence)"
FT                   /evidence="ECO:0000305"
SQ   SEQUENCE   614 AA;  66750 MW;  37B78FEB06A4A3D1 CRC64;
     MQEKTKRIIK EDIEAVRAYS DCFDVEMPDL DENGEVIGLP APYPREVAGT VRSGYRIYDL
     AKKAKERGWP IQNPILGRNT AEETYGESQE MYAYADKFDE TLFHFVHAEA TRHIDPLKGR
     ELINQSRGKG GITPIGEREF IAMGGGSKHP VRINATGDTP HLSIINALIA GFDGTDIGPV
     IHVHFGGRGI HDYKTKVVNG YKAIQICAEN NIFVQLDSHK HLNNIGGTDG MALAMCLLSE
     GLAVHAGLPW ELSAIQMNVA GINIYADLAV MRAFRKACHS KSIIAVPETF QNPPGNLVAE
     AAHFSRMAVT AKLGGADFYR PKAAESVGIP TGDSMGQAIW GTEDVFGHVV NPDIQSPVID
     AREAEIIDEA LAVLEATLHL EGLTLEAMTD DFWKQWSDEA LIDLIVAAGK AGVLDSQRAA
     GWDLKRHVVV NRDKDGITRY VKGYTPLGVD ASRCAQSDED VEVHVEKAPT RPEKIVLATV
     GADAHVNGIN VIREAFQDAG YDVVYLRGMN LPESVAEVAA EVGADAVGVS NLLGLGMELF
     PRVSKRLEEL GLRDKMVVCA GGRIAEKEEE HRQFEEKIQK EGSAFMGMDG FFGPGSSPED
     CVKIIGDMIN AKKA
//