ID MEDH_BACMT Reviewed; 381 AA. AC P31005; DT 01-JUL-1993, integrated into UniProtKB/Swiss-Prot. DT 23-JAN-2007, sequence version 3. DT 02-SEP-2026, entry version 88. DE RecName: Full=NAD-dependent methanol dehydrogenase {ECO:0000303|PubMed:1995642}; DE Short=MDH {ECO:0000303|PubMed:1995642}; DE Short=MEDH {ECO:0000303|PubMed:2673121}; DE EC=1.1.1.244 {ECO:0000269|PubMed:12089158, ECO:0000269|PubMed:12351635, ECO:0000269|PubMed:1995643, ECO:0000269|PubMed:2673121}; DE AltName: Full=Type 3 alcohol dehydrogenase {ECO:0000303|PubMed:1644761}; GN Name=mdh {ECO:0000303|PubMed:2673121}; OS Bacillus methanolicus. OC Bacteria; Bacillati; Bacillota; Bacilli; Caryophanales; Bacillaceae; OC Bacillus. OX NCBI_TaxID=1471; RN [1] RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], PROTEIN SEQUENCE OF 2-15, FUNCTION, RP COFACTOR, AND ACTIVITY REGULATION. RC STRAIN=C1; RX PubMed=1644761; DOI=10.1128/jb.174.16.5346-5353.1992; RA de Vries G.E., Arfman N., Terpstra P., Dijkhuizen L.; RT "Cloning, expression, and sequence analysis of the Bacillus methanolicus C1 RT methanol dehydrogenase gene."; RL J. Bacteriol. 174:5346-5353(1992). RN [2] RP PROTEIN SEQUENCE OF 2-61, FUNCTION, COFACTOR, ACTIVITY REGULATION, AND RP SUBUNIT. RC STRAIN=C1; RX PubMed=1995642; DOI=10.1016/s0021-9258(19)67885-3; RA Vonck J., Arfman N., de Vries G.E., van Beeumen J., van Bruggen E.F.J., RA Dijkhuizen L.; RT "Electron microscopic analysis and biochemical characterization of a novel RT methanol dehydrogenase from the thermotolerant Bacillus sp. C1."; RL J. Biol. Chem. 266:3949-3954(1991). RN [3] RP FUNCTION, CATALYTIC ACTIVITY, SUBCELLULAR LOCATION, SUBUNIT, AND SUBSTRATE RP SPECIFICITY. RC STRAIN=C1; RX PubMed=2673121; DOI=10.1007/bf00409664; RA Arfman N., Watling E.M., Clement W., van Oosterwijk R.J., de Vries G.E., RA Harder W., Attwood M.M., Dijkhuizen L.; RT "Methanol metabolism in thermotolerant methylotrophic Bacillus strains RT involving a novel catabolic NAD-dependent methanol dehydrogenase as a key RT enzyme."; RL Arch. Microbiol. 152:280-288(1989). RN [4] RP FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, AND ACTIVITY RP REGULATION. RC STRAIN=C1; RX PubMed=1995643; DOI=10.1016/s0021-9258(19)67886-5; RA Arfman N., Van Beeumen J., De Vries G.E., Harder W., Dijkhuizen L.; RT "Purification and characterization of an activator protein for methanol RT dehydrogenase from thermotolerant Bacillus spp."; RL J. Biol. Chem. 266:3955-3960(1991). RN [5] RP FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, AND REACTION RP MECHANISM. RC STRAIN=C1; RX PubMed=12089158; DOI=10.1074/jbc.m205617200; RA Kloosterman H., Vrijbloed J.W., Dijkhuizen L.; RT "Molecular, biochemical, and functional characterization of a Nudix RT hydrolase protein that stimulates the activity of a nicotinoprotein alcohol RT dehydrogenase."; RL J. Biol. Chem. 277:34785-34792(2002). RN [6] RP FUNCTION, CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, MUTAGENESIS OF RP GLY-13; GLY-15; ASP-88; GLY-95; SER-97; ASP-100 AND LYS-103, AND COFACTOR. RC STRAIN=C1; RX PubMed=12351635; DOI=10.1074/jbc.m207547200; RA Hektor H.J., Kloosterman H., Dijkhuizen L.; RT "Identification of a magnesium-dependent NAD(P)(H)-binding domain in the RT nicotinoprotein methanol dehydrogenase from Bacillus methanolicus."; RL J. Biol. Chem. 277:46966-46973(2002). CC -!- FUNCTION: Catalyzes the oxidation of methanol to yield formaldehyde. It CC possesses a NADH-dependent formaldehyde reductase activity and cannot CC use NADP. {ECO:0000269|PubMed:12089158, ECO:0000269|PubMed:12351635, CC ECO:0000269|PubMed:1644761, ECO:0000269|PubMed:1995642, CC ECO:0000269|PubMed:1995643, ECO:0000269|PubMed:2673121}. CC -!- CATALYTIC ACTIVITY: CC Reaction=methanol + NAD(+) = formaldehyde + NADH + H(+); CC Xref=Rhea:RHEA:19401, ChEBI:CHEBI:15378, ChEBI:CHEBI:16842, CC ChEBI:CHEBI:17790, ChEBI:CHEBI:57540, ChEBI:CHEBI:57945; CC EC=1.1.1.244; Evidence={ECO:0000269|PubMed:12089158, CC ECO:0000269|PubMed:12351635, ECO:0000269|PubMed:1995643, CC ECO:0000269|PubMed:2673121}; CC -!- COFACTOR: CC Name=Mg(2+); Xref=ChEBI:CHEBI:18420; CC Evidence={ECO:0000269|PubMed:12351635, ECO:0000269|PubMed:1644761, CC ECO:0000269|PubMed:1995642, ECO:0000269|PubMed:2673121}; CC -!- COFACTOR: CC Name=Zn(2+); Xref=ChEBI:CHEBI:29105; CC Evidence={ECO:0000269|PubMed:12351635, ECO:0000269|PubMed:1644761, CC ECO:0000269|PubMed:1995642}; CC -!- ACTIVITY REGULATION: Stimulated by the activator protein Act which CC requires the presence of magnesium ions. Inhibited by 1,10- CC phenanthroline. {ECO:0000269|PubMed:1644761, CC ECO:0000269|PubMed:1995642, ECO:0000269|PubMed:1995643}. CC -!- BIOPHYSICOCHEMICAL PROPERTIES: CC Kinetic parameters: CC KM=9 uM for NADH {ECO:0000269|PubMed:12351635}; CC KM=0.02 mM for NAD {ECO:0000269|PubMed:12089158, CC ECO:0000269|PubMed:12351635}; CC KM=0.7 mM for formaldehyde (without activator protein at pH 9.5 and CC 50 degrees Celsius) {ECO:0000269|PubMed:1995643}; CC KM=2 mM for formaldehyde (with activator protein at pH 9.5 and 50 CC degrees Celsius) {ECO:0000269|PubMed:1995643}; CC KM=57 mM for ethanol (with the activator protein at pH 9.5 and 50 CC degrees Celsius) {ECO:0000269|PubMed:1995643}; CC KM=94 mM for ethanol (without activator protein at pH 9.5 and 50 CC degrees Celsius) {ECO:0000269|PubMed:1995643}; CC KM=140 mM for methanol (with the activator protein at pH 9.5 and 50 CC degrees Celsius) {ECO:0000269|PubMed:1995643}; CC KM=230 mM for methanol (without activator protein at pH 9.5 and 50 CC degrees Celsius) {ECO:0000269|PubMed:1995643}; CC Vmax=1310 umol/min/mg enzyme {ECO:0000269|PubMed:12089158, CC ECO:0000269|PubMed:12351635}; CC -!- PATHWAY: One-carbon metabolism; methanol degradation; formaldehyde from CC methanol: step 1/1. {ECO:0000305}. CC -!- SUBUNIT: Homodecamer. {ECO:0000269|PubMed:1995642, CC ECO:0000269|PubMed:2673121}. CC -!- SUBCELLULAR LOCATION: Cytoplasm {ECO:0000269|PubMed:2673121}. CC -!- SIMILARITY: Belongs to the iron-containing alcohol dehydrogenase CC 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; M65004; AAA22593.1; -; Genomic_DNA. DR RefSeq; WP_274857131.1; NZ_PNFF01000007.1. DR PDB; 8ZWD; X-ray; 3.00 A; A/B/C/D/E/F/G/H/I/J=2-381. DR AlphaFoldDB; P31005; -. DR SMR; P31005; -. DR KEGG; ag:AAA22593; -. DR BioCyc; MetaCyc:MONOMER-15636; -. DR BRENDA; 1.1.1.244; 674. DR SABIO-RK; P31005; -. DR UniPathway; UPA00928; UER00893. DR GO; GO:0005737; C:cytoplasm; IDA:UniProtKB. DR GO; GO:0000287; F:magnesium ion binding; IDA:UniProtKB. DR GO; GO:0050093; F:methanol dehydrogenase (NAD+) activity; IDA:UniProtKB. DR GO; GO:0008270; F:zinc ion binding; IDA:UniProtKB. DR GO; GO:0046170; P:methanol catabolic process; IEA:UniProtKB-UniPathway. DR CDD; cd08188; PDDH; 1. DR FunFam; 3.40.50.1970:FF:000003; Alcohol dehydrogenase, iron-containing; 1. DR FunFam; 1.20.1090.10:FF:000001; Aldehyde-alcohol dehydrogenase; 1. DR Gene3D; 3.40.50.1970; -; 1. DR Gene3D; 1.20.1090.10; Dehydroquinate synthase-like - alpha domain; 1. DR InterPro; IPR001670; ADH_Fe/GldA. DR InterPro; IPR056798; ADH_Fe_C. DR InterPro; IPR018211; ADH_Fe_CS. DR InterPro; IPR039697; Alcohol_dehydrogenase_Fe. DR PANTHER; PTHR11496; ALCOHOL DEHYDROGENASE; 1. DR PANTHER; PTHR11496:SF102; ALCOHOL DEHYDROGENASE 4; 1. DR Pfam; PF25137; ADH_Fe_C; 1. DR Pfam; PF00465; Fe-ADH; 1. DR SUPFAM; SSF56796; Dehydroquinate synthase-like; 1. DR PROSITE; PS00913; ADH_IRON_1; 1. DR PROSITE; PS00060; ADH_IRON_2; 1. DR PDBsum; 8ZWD; -. PE 1: Evidence at protein level; KW 3D-structure; Cytoplasm; Direct protein sequencing; Magnesium; KW Methanol utilization; NAD; Oxidoreductase; Zinc. FT INIT_MET 1 FT /note="Removed" FT /evidence="ECO:0000269|PubMed:1644761, FT ECO:0000269|PubMed:1995642" FT CHAIN 2..381 FT /note="NAD-dependent methanol dehydrogenase" FT /id="PRO_0000087834" FT MUTAGEN 13 FT /note="G->A: Shows a reduced dehydrogenase activity." FT /evidence="ECO:0000269|PubMed:12351635" FT MUTAGEN 15 FT /note="G->A: Shows almost the same dehydrogenase activity FT as the wild-type." FT /evidence="ECO:0000269|PubMed:12351635" FT MUTAGEN 88 FT /note="D->N: Shows almost the same dehydrogenase activity FT as the wild-type." FT /evidence="ECO:0000269|PubMed:12351635" FT MUTAGEN 95 FT /note="G->A: Shows a 10-fold decreased affinity for NAD and FT NADH and a strongly reduced dehydrogenase activity. FT Completely insensitive to the stimulating effect of the FT activator protein Act." FT /evidence="ECO:0000269|PubMed:12351635" FT MUTAGEN 97 FT /note="S->G: Shows an increase of the dehydrogenase FT activity and a decrease of the affinity for NAD and NADH. FT Completely insensitive to the stimulating effect of the FT activator protein Act. It does not bind NAD." FT /evidence="ECO:0000269|PubMed:12351635" FT MUTAGEN 97 FT /note="S->T: Shows an increase of the dehydrogenase FT activity and affinity for NAD and NADH." FT /evidence="ECO:0000269|PubMed:12351635" FT MUTAGEN 100 FT /note="D->N: Loss of dehydrogenase activity. It still binds FT NADH." FT /evidence="ECO:0000269|PubMed:12351635" FT MUTAGEN 103 FT /note="K->R: Loss of dehydrogenase activity. It does not FT bind NADH." FT /evidence="ECO:0000269|PubMed:12351635" FT CONFLICT 18 FT /note="K -> L (in Ref. 2; AA sequence)" FT /evidence="ECO:0000305" FT CONFLICT 42 FT /note="S -> A (in Ref. 2; AA sequence)" FT /evidence="ECO:0000305" FT CONFLICT 50 FT /note="A -> D (in Ref. 2; AA sequence)" FT /evidence="ECO:0000305" FT CONFLICT 59 FT /note="D -> N (in Ref. 2; AA sequence)" FT /evidence="ECO:0000305" FT STRAND 2..5 FT /evidence="ECO:0007829|PDB:8ZWD" FT STRAND 8..13 FT /evidence="ECO:0007829|PDB:8ZWD" FT HELIX 16..19 FT /evidence="ECO:0007829|PDB:8ZWD" FT HELIX 20..26 FT /evidence="ECO:0007829|PDB:8ZWD" FT STRAND 30..35 FT /evidence="ECO:0007829|PDB:8ZWD" FT HELIX 38..43 FT /evidence="ECO:0007829|PDB:8ZWD" FT HELIX 45..55 FT /evidence="ECO:0007829|PDB:8ZWD" FT STRAND 59..63 FT /evidence="ECO:0007829|PDB:8ZWD" FT HELIX 72..85 FT /evidence="ECO:0007829|PDB:8ZWD" FT STRAND 88..95 FT /evidence="ECO:0007829|PDB:8ZWD" FT HELIX 96..111 FT /evidence="ECO:0007829|PDB:8ZWD" FT HELIX 115..118 FT /evidence="ECO:0007829|PDB:8ZWD" FT STRAND 131..135 FT /evidence="ECO:0007829|PDB:8ZWD" FT STRAND 137..139 FT /evidence="ECO:0007829|PDB:8ZWD" FT HELIX 142..144 FT /evidence="ECO:0007829|PDB:8ZWD" FT STRAND 146..152 FT /evidence="ECO:0007829|PDB:8ZWD" FT TURN 153..156 FT /evidence="ECO:0007829|PDB:8ZWD" FT STRAND 157..162 FT /evidence="ECO:0007829|PDB:8ZWD" FT HELIX 164..166 FT /evidence="ECO:0007829|PDB:8ZWD" FT STRAND 169..173 FT /evidence="ECO:0007829|PDB:8ZWD" FT HELIX 175..178 FT /evidence="ECO:0007829|PDB:8ZWD" FT HELIX 183..202 FT /evidence="ECO:0007829|PDB:8ZWD" FT HELIX 208..230 FT /evidence="ECO:0007829|PDB:8ZWD" FT HELIX 235..255 FT /evidence="ECO:0007829|PDB:8ZWD" FT HELIX 259..271 FT /evidence="ECO:0007829|PDB:8ZWD" FT HELIX 275..290 FT /evidence="ECO:0007829|PDB:8ZWD" FT TURN 291..293 FT /evidence="ECO:0007829|PDB:8ZWD" FT HELIX 295..304 FT /evidence="ECO:0007829|PDB:8ZWD" FT HELIX 314..331 FT /evidence="ECO:0007829|PDB:8ZWD" FT TURN 338..342 FT /evidence="ECO:0007829|PDB:8ZWD" FT HELIX 345..347 FT /evidence="ECO:0007829|PDB:8ZWD" FT HELIX 348..356 FT /evidence="ECO:0007829|PDB:8ZWD" FT HELIX 360..363 FT /evidence="ECO:0007829|PDB:8ZWD" FT STRAND 364..366 FT /evidence="ECO:0007829|PDB:8ZWD" FT HELIX 370..379 FT /evidence="ECO:0007829|PDB:8ZWD" SQ SEQUENCE 381 AA; 40045 MW; 8EA251B36F73749D CRC64; MTNFFIPPAS VIGRGAVKEV GTRLKQIGAK KALIVTDAFL HSTGLSEEVA KNIREAGLDV AIFPKAQPDP ADTQVHEGVD VFKQENCDAL VSIGGGSSHD TAKAIGLVAA NGGRINDYQG VNSVEKPVVP VVAITTTAGT GSETTSLAVI TDSARKVKMP VIDEKITPTV AIVDPELMVK KPAGLTIATG MDALSHAIEA YVAKGATPVT DAFAIQAMKL INEYLPKAVA NGEDIEAREA MAYAQYMAGV AFNNGGLGLV HSISHQVGGV YKLQHGICNS VNMPHVCAFN LIAKTERFAH IAELLGENVS GLSTAAAAER AIVALERYNK NFGIPSGYAE MGVKEEDIEL LAKNAFEDVC TQSNPRVATV QDIAQIIKNA L //