ID DMGO_ARTGO Reviewed; 830 AA. AC Q9AGP8; DT 14-MAY-2014, integrated into UniProtKB/Swiss-Prot. DT 01-JUN-2001, sequence version 1. DT 27-MAR-2024, entry version 104. DE RecName: Full=Dimethylglycine oxidase; DE Short=DMGO; DE EC=1.5.3.10; GN Name=dmg; OS Arthrobacter globiformis. OC Bacteria; Actinomycetota; Actinomycetes; Micrococcales; Micrococcaceae; OC Arthrobacter. OX NCBI_TaxID=1665; RN [1] RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], FUNCTION, CATALYTIC ACTIVITY, COFACTOR, RP AND BIOPHYSICOCHEMICAL PROPERTIES. RC STRAIN=NRRL B-2979; RX PubMed=11422368; DOI=10.1046/j.1432-1327.2001.02239.x; RA Meskys R., Harris R.J., Casaite V., Basran J., Scrutton N.S.; RT "Organization of the genes involved in dimethylglycine and sarcosine RT degradation in Arthrobacter spp.: implications for glycine betaine RT catabolism."; RL Eur. J. Biochem. 268:3390-3398(2001). RN [2] RP PROTEIN SEQUENCE OF 27-58, FUNCTION, AND FAD-BINDING AT HIS-48. RC STRAIN=NRRL B-2979; RX PubMed=11926836; DOI=10.1021/bi025519h; RA Basran J., Bhanji N., Basran A., Nietlispach D., Mistry S., Meskys R., RA Scrutton N.S.; RT "Mechanistic aspects of the covalent flavoprotein dimethylglycine oxidase RT of Arthrobacter globiformis studied by stopped-flow spectrophotometry."; RL Biochemistry 41:4733-4743(2002). RN [3] RP X-RAY CRYSTALLOGRAPHY (1.61 ANGSTROMS) IN COMPLEX WITH FAD AND RP TETRAHYDROFOLATE, FUNCTION, AND MUTAGENESIS OF HIS-225 AND TYR-259. RC STRAIN=NRRL B-2979; RX PubMed=12912903; DOI=10.1093/emboj/cdg395; RA Leys D., Basran J., Scrutton N.S.; RT "Channelling and formation of 'active' formaldehyde in dimethylglycine RT oxidase."; RL EMBO J. 22:4038-4048(2003). RN [4] RP X-RAY CRYSTALLOGRAPHY (2.00 ANGSTROMS) OF 4-830 IN COMPLEX WITH FAD AND RP TETRAHYDROFOLATE, FUNCTION, AND MUTAGENESIS OF ASP-552. RC STRAIN=NRRL B-2979; RX PubMed=19369258; DOI=10.1074/jbc.m109.006262; RA Tralau T., Lafite P., Levy C., Combe J.P., Scrutton N.S., Leys D.; RT "An internal reaction chamber in dimethylglycine oxidase provides efficient RT protection from exposure to toxic formaldehyde."; RL J. Biol. Chem. 284:17826-17834(2009). CC -!- FUNCTION: Catalyzes the oxidative demethylation of N,N-dimethylglycine CC to yield sarcosine, formaldehyde and hydrogen peroxide. The oxidation CC of dimethylglycine is coupled to the synthesis of 5,10- CC methylenetetrahydrofolate through an unusual substrate channeling CC mechanism. This channeling occurs by nonbiased diffusion of the iminium CC intermediate through a large solvent cavity connecting active site 1 CC (N-terminus) and active site 2 (C-terminus). The synthesis of 5,10- CC methylenetetrahydrofolate (at active site 2) prevents the accumulation CC of formaldehyde, formed by hydrolysis of the iminium intermediate CC product (at active site 1). Does not oxidize sarcosine. CC {ECO:0000269|PubMed:11422368, ECO:0000269|PubMed:11926836, CC ECO:0000269|PubMed:12912903, ECO:0000269|PubMed:19369258}. CC -!- CATALYTIC ACTIVITY: CC Reaction=H2O + N,N-dimethylglycine + O2 = formaldehyde + H2O2 + CC sarcosine; Xref=Rhea:RHEA:17077, ChEBI:CHEBI:15377, CC ChEBI:CHEBI:15379, ChEBI:CHEBI:16240, ChEBI:CHEBI:16842, CC ChEBI:CHEBI:57433, ChEBI:CHEBI:58251; EC=1.5.3.10; CC Evidence={ECO:0000269|PubMed:11422368}; CC -!- CATALYTIC ACTIVITY: CC Reaction=(6S)-5,6,7,8-tetrahydrofolate + N,N-dimethylglycine + O2 = CC (6R)-5,10-methylene-5,6,7,8-tetrahydrofolate + H2O2 + sarcosine; CC Xref=Rhea:RHEA:45756, ChEBI:CHEBI:15379, ChEBI:CHEBI:15636, CC ChEBI:CHEBI:16240, ChEBI:CHEBI:57433, ChEBI:CHEBI:57453, CC ChEBI:CHEBI:58251; Evidence={ECO:0000269|PubMed:11422368}; CC -!- COFACTOR: CC Name=FAD; Xref=ChEBI:CHEBI:57692; CC Evidence={ECO:0000269|PubMed:11422368, ECO:0000269|PubMed:11926836}; CC Note=Binds 1 FAD per subunit. {ECO:0000269|PubMed:11422368, CC ECO:0000269|PubMed:11926836}; CC -!- BIOPHYSICOCHEMICAL PROPERTIES: CC Kinetic parameters: CC KM=2 mM for N,N-dimethylglycine {ECO:0000269|PubMed:11422368}; CC pH dependence: CC Optimum pH is 8.0-10.0. {ECO:0000269|PubMed:11422368}; CC -!- SIMILARITY: Belongs to the GcvT family. {ECO:0000305}. 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; AF329477; AAK16482.1; -; Genomic_DNA. DR PDB; 1PJ5; X-ray; 1.61 A; A=1-830. DR PDB; 1PJ6; X-ray; 1.65 A; A=1-830. DR PDB; 1PJ7; X-ray; 2.10 A; A=1-830. DR PDB; 3GSI; X-ray; 2.00 A; A=4-830. DR PDBsum; 1PJ5; -. DR PDBsum; 1PJ6; -. DR PDBsum; 1PJ7; -. DR PDBsum; 3GSI; -. DR AlphaFoldDB; Q9AGP8; -. DR SMR; Q9AGP8; -. DR DrugBank; DB03256; (6R)-Folinic acid. DR DrugBank; DB03147; Flavin adenine dinucleotide. DR KEGG; ag:AAK16482; -. DR BioCyc; MetaCyc:MONOMER-21836; -. DR BRENDA; 1.5.3.10; 444. DR SABIO-RK; Q9AGP8; -. DR EvolutionaryTrace; Q9AGP8; -. DR GO; GO:0047866; F:dimethylglycine oxidase activity; IEA:UniProtKB-EC. DR GO; GO:0000166; F:nucleotide binding; IEA:UniProtKB-KW. DR Gene3D; 2.40.30.110; Aminomethyltransferase beta-barrel domains; 1. DR Gene3D; 3.30.70.1400; Aminomethyltransferase beta-barrel domains; 1. DR Gene3D; 3.30.9.10; D-Amino Acid Oxidase, subunit A, domain 2; 1. DR Gene3D; 3.50.50.60; FAD/NAD(P)-binding domain; 1. DR InterPro; IPR006076; FAD-dep_OxRdtase. DR InterPro; IPR036188; FAD/NAD-bd_sf. DR InterPro; IPR032503; FAO_M. DR InterPro; IPR028896; GCST/YgfZ/DmdA. DR InterPro; IPR013977; GCV_T_C. DR InterPro; IPR006222; GCV_T_N. DR InterPro; IPR029043; GcvT/YgfZ_C. DR InterPro; IPR027266; TrmE/GcvT_dom1. DR PANTHER; PTHR43757; AMINOMETHYLTRANSFERASE; 1. DR PANTHER; PTHR43757:SF2; AMINOMETHYLTRANSFERASE, MITOCHONDRIAL; 1. DR Pfam; PF01266; DAO; 1. DR Pfam; PF16350; FAO_M; 1. DR Pfam; PF01571; GCV_T; 1. DR Pfam; PF08669; GCV_T_C; 1. DR SUPFAM; SSF101790; Aminomethyltransferase beta-barrel domain; 1. DR SUPFAM; SSF54373; FAD-linked reductases, C-terminal domain; 1. DR SUPFAM; SSF51905; FAD/NAD(P)-binding domain; 1. DR SUPFAM; SSF103025; Folate-binding domain; 1. PE 1: Evidence at protein level; KW 3D-structure; Direct protein sequencing; FAD; Flavoprotein; KW Nucleotide-binding; Oxidoreductase. FT CHAIN 1..830 FT /note="Dimethylglycine oxidase" FT /id="PRO_0000428956" FT ACT_SITE 225 FT /evidence="ECO:0000255" FT ACT_SITE 259 FT /evidence="ECO:0000255" FT ACT_SITE 552 FT /note="For 5,10-methylenetetrahydrofolate synthesis FT activity" FT /evidence="ECO:0000255" FT BINDING 14..15 FT /ligand="FAD" FT /ligand_id="ChEBI:CHEBI:57692" FT /evidence="ECO:0000269|PubMed:12912903, FT ECO:0000269|PubMed:19369258" FT BINDING 35..36 FT /ligand="FAD" FT /ligand_id="ChEBI:CHEBI:57692" FT /evidence="ECO:0000269|PubMed:12912903, FT ECO:0000269|PubMed:19369258" FT BINDING 45..48 FT /ligand="FAD" FT /ligand_id="ChEBI:CHEBI:57692" FT /evidence="ECO:0000269|PubMed:12912903, FT ECO:0000269|PubMed:19369258" FT BINDING 52 FT /ligand="FAD" FT /ligand_id="ChEBI:CHEBI:57692" FT /evidence="ECO:0000269|PubMed:12912903, FT ECO:0000269|PubMed:19369258" FT BINDING 174 FT /ligand="FAD" FT /ligand_id="ChEBI:CHEBI:57692" FT /evidence="ECO:0000269|PubMed:12912903, FT ECO:0000269|PubMed:19369258" FT BINDING 259 FT /ligand="FAD" FT /ligand_id="ChEBI:CHEBI:57692" FT /evidence="ECO:0000269|PubMed:12912903, FT ECO:0000269|PubMed:19369258" FT BINDING 360..363 FT /ligand="FAD" FT /ligand_id="ChEBI:CHEBI:57692" FT /evidence="ECO:0000269|PubMed:12912903, FT ECO:0000269|PubMed:19369258" FT BINDING 539 FT /ligand="(6S)-5,6,7,8-tetrahydrofolate" FT /ligand_id="ChEBI:CHEBI:57453" FT /evidence="ECO:0000269|PubMed:19369258" FT BINDING 554 FT /ligand="(6S)-5,6,7,8-tetrahydrofolate" FT /ligand_id="ChEBI:CHEBI:57453" FT BINDING 566 FT /ligand="(6S)-5,6,7,8-tetrahydrofolate" FT /ligand_id="ChEBI:CHEBI:57453" FT BINDING 658..660 FT /ligand="(6S)-5,6,7,8-tetrahydrofolate" FT /ligand_id="ChEBI:CHEBI:57453" FT SITE 225 FT /note="Important for catalytic activity" FT /evidence="ECO:0000269|PubMed:12912903" FT SITE 259 FT /note="Important for catalytic activity" FT /evidence="ECO:0000269|PubMed:12912903" FT SITE 552 FT /note="Important for catalytic activity" FT /evidence="ECO:0000269|PubMed:19369258, FT ECO:0000305|PubMed:12912903" FT MOD_RES 48 FT /note="Pros-8alpha-FAD histidine" FT MUTAGEN 225 FT /note="H->Q: Reduces catalytic efficiency 3-fold and FT substrate affinity 30-fold." FT /evidence="ECO:0000269|PubMed:12912903" FT MUTAGEN 259 FT /note="Y->F: Reduces catalytic efficiency 225-fold and FT substrate affinity 25-fold." FT /evidence="ECO:0000269|PubMed:12912903" FT MUTAGEN 552 FT /note="D->A: No effect on the activity." FT /evidence="ECO:0000269|PubMed:19369258" FT MUTAGEN 552 FT /note="D->N: Reduces activity 3-fold." FT /evidence="ECO:0000269|PubMed:19369258" FT STRAND 7..10 FT /evidence="ECO:0007829|PDB:1PJ5" FT HELIX 14..25 FT /evidence="ECO:0007829|PDB:1PJ5" FT STRAND 31..34 FT /evidence="ECO:0007829|PDB:1PJ5" FT HELIX 45..47 FT /evidence="ECO:0007829|PDB:1PJ5" FT HELIX 60..75 FT /evidence="ECO:0007829|PDB:1PJ5" FT STRAND 81..85 FT /evidence="ECO:0007829|PDB:1PJ5" FT STRAND 89..95 FT /evidence="ECO:0007829|PDB:1PJ5" FT HELIX 96..112 FT /evidence="ECO:0007829|PDB:1PJ5" FT STRAND 117..119 FT /evidence="ECO:0007829|PDB:1PJ5" FT HELIX 121..127 FT /evidence="ECO:0007829|PDB:1PJ5" FT HELIX 133..135 FT /evidence="ECO:0007829|PDB:1PJ5" FT STRAND 138..142 FT /evidence="ECO:0007829|PDB:1PJ5" FT STRAND 146..148 FT /evidence="ECO:0007829|PDB:1PJ5" FT HELIX 150..163 FT /evidence="ECO:0007829|PDB:1PJ5" FT STRAND 167..169 FT /evidence="ECO:0007829|PDB:1PJ5" FT STRAND 174..180 FT /evidence="ECO:0007829|PDB:1PJ5" FT STRAND 183..189 FT /evidence="ECO:0007829|PDB:1PJ5" FT STRAND 192..195 FT /evidence="ECO:0007829|PDB:1PJ5" FT STRAND 197..201 FT /evidence="ECO:0007829|PDB:1PJ5" FT HELIX 204..206 FT /evidence="ECO:0007829|PDB:1PJ5" FT HELIX 207..212 FT /evidence="ECO:0007829|PDB:1PJ5" FT TURN 213..215 FT /evidence="ECO:0007829|PDB:1PJ5" FT STRAND 221..231 FT /evidence="ECO:0007829|PDB:1PJ5" FT HELIX 234..236 FT /evidence="ECO:0007829|PDB:1PJ5" FT TURN 237..239 FT /evidence="ECO:0007829|PDB:1PJ5" FT TURN 242..244 FT /evidence="ECO:0007829|PDB:1PJ5" FT STRAND 250..253 FT /evidence="ECO:0007829|PDB:1PJ5" FT HELIX 254..256 FT /evidence="ECO:0007829|PDB:1PJ5" FT STRAND 258..263 FT /evidence="ECO:0007829|PDB:1PJ5" FT STRAND 266..271 FT /evidence="ECO:0007829|PDB:1PJ5" FT HELIX 281..283 FT /evidence="ECO:0007829|PDB:1PJ5" FT HELIX 289..291 FT /evidence="ECO:0007829|PDB:1PJ5" FT HELIX 305..318 FT /evidence="ECO:0007829|PDB:1PJ5" FT HELIX 320..324 FT /evidence="ECO:0007829|PDB:1PJ5" FT STRAND 327..337 FT /evidence="ECO:0007829|PDB:1PJ5" FT STRAND 344..347 FT /evidence="ECO:0007829|PDB:1PJ5" FT STRAND 349..352 FT /evidence="ECO:0007829|PDB:1PJ5" FT STRAND 354..359 FT /evidence="ECO:0007829|PDB:1PJ5" FT HELIX 362..364 FT /evidence="ECO:0007829|PDB:1PJ5" FT HELIX 365..378 FT /evidence="ECO:0007829|PDB:1PJ5" FT TURN 386..388 FT /evidence="ECO:0007829|PDB:1PJ5" FT HELIX 390..392 FT /evidence="ECO:0007829|PDB:1PJ5" FT HELIX 395..398 FT /evidence="ECO:0007829|PDB:1PJ5" FT HELIX 400..412 FT /evidence="ECO:0007829|PDB:1PJ5" FT TURN 413..415 FT /evidence="ECO:0007829|PDB:1PJ5" FT STRAND 425..427 FT /evidence="ECO:0007829|PDB:1PJ5" FT HELIX 436..441 FT /evidence="ECO:0007829|PDB:1PJ5" FT STRAND 444..449 FT /evidence="ECO:0007829|PDB:1PJ5" FT STRAND 452..458 FT /evidence="ECO:0007829|PDB:1PJ5" FT HELIX 459..467 FT /evidence="ECO:0007829|PDB:1PJ5" FT HELIX 470..472 FT /evidence="ECO:0007829|PDB:1PJ5" FT HELIX 479..482 FT /evidence="ECO:0007829|PDB:1PJ5" FT HELIX 488..498 FT /evidence="ECO:0007829|PDB:1PJ5" FT STRAND 501..504 FT /evidence="ECO:0007829|PDB:1PJ5" FT STRAND 510..515 FT /evidence="ECO:0007829|PDB:1PJ5" FT HELIX 518..525 FT /evidence="ECO:0007829|PDB:1PJ5" FT STRAND 526..528 FT /evidence="ECO:0007829|PDB:1PJ5" FT STRAND 536..543 FT /evidence="ECO:0007829|PDB:1PJ5" FT STRAND 549..559 FT /evidence="ECO:0007829|PDB:1PJ5" FT STRAND 562..566 FT /evidence="ECO:0007829|PDB:1PJ5" FT HELIX 570..586 FT /evidence="ECO:0007829|PDB:1PJ5" FT STRAND 594..597 FT /evidence="ECO:0007829|PDB:1PJ5" FT HELIX 599..601 FT /evidence="ECO:0007829|PDB:1PJ5" FT STRAND 602..609 FT /evidence="ECO:0007829|PDB:1PJ5" FT HELIX 612..616 FT /evidence="ECO:0007829|PDB:1PJ5" FT TURN 617..619 FT /evidence="ECO:0007829|PDB:1PJ5" FT TURN 626..628 FT /evidence="ECO:0007829|PDB:1PJ5" FT STRAND 633..639 FT /evidence="ECO:0007829|PDB:1PJ5" FT STRAND 642..647 FT /evidence="ECO:0007829|PDB:1PJ5" FT STRAND 653..662 FT /evidence="ECO:0007829|PDB:1PJ5" FT HELIX 663..677 FT /evidence="ECO:0007829|PDB:1PJ5" FT HELIX 678..680 FT /evidence="ECO:0007829|PDB:1PJ5" FT STRAND 683..685 FT /evidence="ECO:0007829|PDB:1PJ5" FT HELIX 687..696 FT /evidence="ECO:0007829|PDB:1PJ5" FT TURN 702..704 FT /evidence="ECO:0007829|PDB:1PJ5" FT TURN 712..716 FT /evidence="ECO:0007829|PDB:1PJ5" FT HELIX 718..720 FT /evidence="ECO:0007829|PDB:1PJ5" FT HELIX 731..734 FT /evidence="ECO:0007829|PDB:1PJ5" FT TURN 739..741 FT /evidence="ECO:0007829|PDB:3GSI" FT STRAND 743..751 FT /evidence="ECO:0007829|PDB:1PJ5" FT STRAND 763..766 FT /evidence="ECO:0007829|PDB:1PJ5" FT STRAND 769..773 FT /evidence="ECO:0007829|PDB:1PJ5" FT STRAND 776..780 FT /evidence="ECO:0007829|PDB:1PJ5" FT TURN 781..784 FT /evidence="ECO:0007829|PDB:1PJ5" FT STRAND 785..793 FT /evidence="ECO:0007829|PDB:1PJ5" FT STRAND 801..806 FT /evidence="ECO:0007829|PDB:1PJ5" FT STRAND 809..816 FT /evidence="ECO:0007829|PDB:1PJ5" FT TURN 826..828 FT /evidence="ECO:0007829|PDB:1PJ5" SQ SEQUENCE 830 AA; 89985 MW; 13DE3C4B3DF325DA CRC64; MASTPRIVII GAGIVGTNLA DELVTRGWNN ITVLDQGPLN MPGGSTSHAP GLVFQTNPSK TMASFAKYTV EKLLSLTEDG VSCFNQVGGL EVATTETRLA DLKRKLGYAA AWGIEGRLLS PAECQELYPL LDGENILGGL HVPSDGLASA ARAVQLLIKR TESAGVTYRG STTVTGIEQS GGRVTGVQTA DGVIPADIVV SCAGFWGAKI GAMIGMAVPL LPLAHQYVKT TPVPAQQGRN DQPNGARLPI LRHQDQDLYY REHGDRYGIG SYAHRPMPVD VDTLGAYAPE TVSEHHMPSR LDFTLEDFLP AWEATKQLLP ALADSEIEDG FNGIFSFTPD GGPLLGESKE LDGFYVAEAV WVTHSAGVAK AMAELLTTGR SETDLGECDI TRFEDVQLTP EYVSETSQQN FVEIYDVLHP LQPRLSPRNL RVSPFHARHK ELGAFFLEAG GWERPYWFEA NAALLKEMPA EWLPPARDAW SGMFSSPIAA AEAWKTRTAV AMYDMTPLKR LEVSGPGALK LLQELTTADL AKKPGAVTYT LLLDHAGGVR SDITVARLSE DTFQLGANGN IDTAYFERAA RHQTQSGSAT DWVQVRDTTG GTCCIGLWGP LARDLVSKVS DDDFTNDGLK YFRAKNVVIG GIPVTAMRLS YVGELGWELY TSADNGQRLW DALWQAGQPF GVIAAGRAAF SSLRLEKGYR SWGTDMTTEH DPFEAGLGFA VKMAKESFIG KGALEGRTEE ASARRLRCLT IDDGRSIVLG KEPVFYKEQA VGYVTSAAYG YTVAKPIAYS YLPGTVSVGD SVDIEYFGRR ITATVTEDPL YDPKMTRLRG //