ID MTDC_MOUSE Reviewed; 350 AA. AC P18155; Q3TMN4; DT 01-NOV-1990, integrated into UniProtKB/Swiss-Prot. DT 01-NOV-1990, sequence version 1. DT 02-SEP-2026, entry version 201. DE RecName: Full=Bifunctional methylenetetrahydrofolate dehydrogenase/cyclohydrolase, mitochondrial; DE Includes: DE RecName: Full=NAD-dependent methylenetetrahydrofolate dehydrogenase; DE EC=1.5.1.15 {ECO:0000269|PubMed:2647744, ECO:0000269|PubMed:3487546}; DE Includes: DE RecName: Full=Methenyltetrahydrofolate cyclohydrolase; DE EC=3.5.4.9 {ECO:0000269|PubMed:2647744, ECO:0000269|PubMed:3487546}; DE Flags: Precursor; GN Name=Mthfd2; Synonyms=Nmdmc; OS Mus musculus (Mouse). OC Eukaryota; Metazoa; Chordata; Craniata; Vertebrata; Euteleostomi; Mammalia; OC Eutheria; Euarchontoglires; Glires; Rodentia; Myomorpha; Muroidea; Muridae; OC Murinae; Mus; Mus. OX NCBI_TaxID=10090; RN [1] RP NUCLEOTIDE SEQUENCE [MRNA], PROTEIN SEQUENCE OF 36-55, AND CATALYTIC RP ACTIVITY. RX PubMed=2647744; DOI=10.1016/s0021-9258(18)83667-5; RA Belanger C., Mackenzie R.E.; RT "Isolation and characterization of cDNA clones encoding the murine NAD- RT dependent methylenetetrahydrofolate dehydrogenase-methenyltetrahydrofolate RT cyclohydrolase."; RL J. Biol. Chem. 264:4837-4843(1989). RN [2] RP NUCLEOTIDE SEQUENCE [GENOMIC DNA]. RX PubMed=1999293; DOI=10.1016/0378-1119(91)90064-i; RA Belanger C., Mackenzie R.E.; RT "Structural organization of the murine gene encoding NAD-dependent RT methylenetetrahydrofolate dehydrogenase-methenyltetrahydrofolate RT cyclohydrolase."; RL Gene 97:283-288(1991). RN [3] RP NUCLEOTIDE SEQUENCE [LARGE SCALE MRNA]. RC STRAIN=C57BL/6J; RX PubMed=16141072; DOI=10.1126/science.1112014; RA Carninci P., Kasukawa T., Katayama S., Gough J., Frith M.C., Maeda N., RA Oyama R., Ravasi T., Lenhard B., Wells C., Kodzius R., Shimokawa K., RA Bajic V.B., Brenner S.E., Batalov S., Forrest A.R., Zavolan M., Davis M.J., RA Wilming L.G., Aidinis V., Allen J.E., Ambesi-Impiombato A., Apweiler R., RA Aturaliya R.N., Bailey T.L., Bansal M., Baxter L., Beisel K.W., Bersano T., RA Bono H., Chalk A.M., Chiu K.P., Choudhary V., Christoffels A., RA Clutterbuck D.R., Crowe M.L., Dalla E., Dalrymple B.P., de Bono B., RA Della Gatta G., di Bernardo D., Down T., Engstrom P., Fagiolini M., RA Faulkner G., Fletcher C.F., Fukushima T., Furuno M., Futaki S., RA Gariboldi M., Georgii-Hemming P., Gingeras T.R., Gojobori T., Green R.E., RA Gustincich S., Harbers M., Hayashi Y., Hensch T.K., Hirokawa N., Hill D., RA Huminiecki L., Iacono M., Ikeo K., Iwama A., Ishikawa T., Jakt M., RA Kanapin A., Katoh M., Kawasawa Y., Kelso J., Kitamura H., Kitano H., RA Kollias G., Krishnan S.P., Kruger A., Kummerfeld S.K., Kurochkin I.V., RA Lareau L.F., Lazarevic D., Lipovich L., Liu J., Liuni S., McWilliam S., RA Madan Babu M., Madera M., Marchionni L., Matsuda H., Matsuzawa S., Miki H., RA Mignone F., Miyake S., Morris K., Mottagui-Tabar S., Mulder N., Nakano N., RA Nakauchi H., Ng P., Nilsson R., Nishiguchi S., Nishikawa S., Nori F., RA Ohara O., Okazaki Y., Orlando V., Pang K.C., Pavan W.J., Pavesi G., RA Pesole G., Petrovsky N., Piazza S., Reed J., Reid J.F., Ring B.Z., RA Ringwald M., Rost B., Ruan Y., Salzberg S.L., Sandelin A., Schneider C., RA Schoenbach C., Sekiguchi K., Semple C.A., Seno S., Sessa L., Sheng Y., RA Shibata Y., Shimada H., Shimada K., Silva D., Sinclair B., Sperling S., RA Stupka E., Sugiura K., Sultana R., Takenaka Y., Taki K., Tammoja K., RA Tan S.L., Tang S., Taylor M.S., Tegner J., Teichmann S.A., Ueda H.R., RA van Nimwegen E., Verardo R., Wei C.L., Yagi K., Yamanishi H., RA Zabarovsky E., Zhu S., Zimmer A., Hide W., Bult C., Grimmond S.M., RA Teasdale R.D., Liu E.T., Brusic V., Quackenbush J., Wahlestedt C., RA Mattick J.S., Hume D.A., Kai C., Sasaki D., Tomaru Y., Fukuda S., RA Kanamori-Katayama M., Suzuki M., Aoki J., Arakawa T., Iida J., Imamura K., RA Itoh M., Kato T., Kawaji H., Kawagashira N., Kawashima T., Kojima M., RA Kondo S., Konno H., Nakano K., Ninomiya N., Nishio T., Okada M., Plessy C., RA Shibata K., Shiraki T., Suzuki S., Tagami M., Waki K., Watahiki A., RA Okamura-Oho Y., Suzuki H., Kawai J., Hayashizaki Y.; RT "The transcriptional landscape of the mammalian genome."; RL Science 309:1559-1563(2005). RN [4] RP NUCLEOTIDE SEQUENCE [LARGE SCALE MRNA]. RC STRAIN=FVB/N; TISSUE=Salivary gland; RX PubMed=15489334; DOI=10.1101/gr.2596504; RG The MGC Project Team; RT "The status, quality, and expansion of the NIH full-length cDNA project: RT the Mammalian Gene Collection (MGC)."; RL Genome Res. 14:2121-2127(2004). RN [5] RP NUCLEOTIDE SEQUENCE [GENOMIC DNA] OF 1-34. RX PubMed=1843253; DOI=10.1093/nar/19.16.4341; RA Belanger C., Peri K.G., Mackenzie R.E.; RT "Analysis of the promoter region of the gene encoding NAD-dependent RT methylenetetrahydrofolate dehydrogenase-methenyltetrahydrofolate RT cyclohydrolase."; RL Nucleic Acids Res. 19:4341-4345(1991). RN [6] RP IDENTIFICATION, CATALYTIC ACTIVITY, AND COFACTOR. RX PubMed=3487546; DOI=10.1016/s0021-9258(18)67686-0; RA Mejia N.R., Rios-Orlandi E.M., MacKenzie R.E.; RT "NAD-dependent methylenetetrahydrofolate dehydrogenase- RT methenyltetrahydrofolate cyclohydrolase from ascites tumor cells. RT Purification and properties."; RL J. Biol. Chem. 261:9509-9513(1986). RN [7] RP IDENTIFICATION BY MASS SPECTROMETRY [LARGE SCALE ANALYSIS]. RC TISSUE=Pancreas, Spleen, and Testis; RX PubMed=21183079; DOI=10.1016/j.cell.2010.12.001; RA Huttlin E.L., Jedrychowski M.P., Elias J.E., Goswami T., Rad R., RA Beausoleil S.A., Villen J., Haas W., Sowa M.E., Gygi S.P.; RT "A tissue-specific atlas of mouse protein phosphorylation and expression."; RL Cell 143:1174-1189(2010). CC -!- FUNCTION: Although its dehydrogenase activity is NAD-specific, it can CC also utilize NADP at a reduced efficiency. CC {ECO:0000250|UniProtKB:P13995}. CC -!- CATALYTIC ACTIVITY: CC Reaction=(6R)-5,10-methylene-5,6,7,8-tetrahydrofolate + NAD(+) = (6R)- CC 5,10-methenyltetrahydrofolate + NADH; Xref=Rhea:RHEA:22892, CC ChEBI:CHEBI:15636, ChEBI:CHEBI:57455, ChEBI:CHEBI:57540, CC ChEBI:CHEBI:57945; EC=1.5.1.15; Evidence={ECO:0000269|PubMed:2647744, CC ECO:0000269|PubMed:3487546}; CC -!- CATALYTIC ACTIVITY: CC Reaction=(6R)-5,10-methenyltetrahydrofolate + H2O = (6R)-10- CC formyltetrahydrofolate + H(+); Xref=Rhea:RHEA:23700, CC ChEBI:CHEBI:15377, ChEBI:CHEBI:15378, ChEBI:CHEBI:57455, CC ChEBI:CHEBI:195366; EC=3.5.4.9; Evidence={ECO:0000269|PubMed:2647744, CC ECO:0000269|PubMed:3487546}; CC -!- COFACTOR: CC Name=Mg(2+); Xref=ChEBI:CHEBI:18420; CC Evidence={ECO:0000269|PubMed:3487546}; CC -!- SUBUNIT: Homodimer. {ECO:0000250|UniProtKB:P13995}. CC -!- SUBCELLULAR LOCATION: Mitochondrion. CC -!- MISCELLANEOUS: This NAD-dependent bifunctional enzyme has very CC different kinetic properties than the larger NADP-dependent CC trifunctional enzyme and is unique in that it requires formation of an CC enzyme-magnesium complex to allow binding of NAD. CC -!- SIMILARITY: Belongs to the tetrahydrofolate CC dehydrogenase/cyclohydrolase 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; J04627; AAA39827.1; -; mRNA. DR EMBL; M63445; AAA39828.1; -; Genomic_DNA. DR EMBL; M63415; AAA39828.1; JOINED; Genomic_DNA. DR EMBL; M63439; AAA39828.1; JOINED; Genomic_DNA. DR EMBL; M63440; AAA39828.1; JOINED; Genomic_DNA. DR EMBL; M63441; AAA39828.1; JOINED; Genomic_DNA. DR EMBL; M63442; AAA39828.1; JOINED; Genomic_DNA. DR EMBL; M63443; AAA39828.1; JOINED; Genomic_DNA. DR EMBL; M63444; AAA39828.1; JOINED; Genomic_DNA. DR EMBL; AK076019; BAC36124.1; -; mRNA. DR EMBL; AK159680; BAE35283.1; -; mRNA. DR EMBL; AK165840; BAE38407.1; -; mRNA. DR EMBL; BC019511; AAH19511.1; -; mRNA. DR EMBL; S52980; -; NOT_ANNOTATED_CDS; Genomic_DNA. DR CCDS; CCDS20277.1; -. DR PIR; A33267; A33267. DR RefSeq; NP_032664.1; NM_008638.2. DR AlphaFoldDB; P18155; -. DR SMR; P18155; -. DR BioGRID; 201594; 4. DR FunCoup; P18155; 1298. DR STRING; 10090.ENSMUSP00000005810; -. DR iPTMnet; P18155; -. DR PhosphoSitePlus; P18155; -. DR PaxDb; 10090-ENSMUSP00000005810; -. DR PeptideAtlas; P18155; -. DR ProteomicsDB; 290108; -. DR Pumba; P18155; -. DR Antibodypedia; 3292; 216 antibodies from 32 providers. DR DNASU; 17768; -. DR Ensembl; ENSMUST00000005810.9; ENSMUSP00000005810.7; ENSMUSG00000005667.9. DR GeneID; 17768; -. DR KEGG; mmu:17768; -. DR UCSC; uc009cne.2; mouse. DR AGR; MGI:1338850; -. DR CTD; 10797; -. DR MGI; MGI:1338850; Mthfd2. DR VEuPathDB; HostDB:ENSMUSG00000005667; -. DR eggNOG; KOG0089; Eukaryota. DR GeneTree; ENSGT00940000154863; -. DR HOGENOM; CLU_034045_0_1_1; -. DR InParanoid; P18155; -. DR OMA; VCHILTK; -. DR OrthoDB; 5126881at2759; -. DR PhylomeDB; P18155; -. DR BRENDA; 1.5.1.15; 3474. DR BRENDA; 3.5.4.9; 3474. DR Reactome; R-MMU-196757; Metabolism of folate and pterines. DR SABIO-RK; P18155; -. DR BioGRID-ORCS; 17768; 9 hits in 79 CRISPR screens. DR PRO; PR:P18155; -. DR Proteomes; UP000000589; Chromosome 6. DR RNAct; P18155; protein. DR Bgee; ENSMUSG00000005667; Expressed in lacrimal gland and 250 other cell types or tissues. DR ExpressionAtlas; P18155; baseline and differential. DR GO; GO:0005759; C:mitochondrial matrix; IDA:MGI. DR GO; GO:0005739; C:mitochondrion; HDA:MGI. DR GO; GO:0000287; F:magnesium ion binding; ISS:UniProtKB. DR GO; GO:0004477; F:methenyltetrahydrofolate cyclohydrolase activity; IDA:UniProtKB. DR GO; GO:0004487; F:methylenetetrahydrofolate dehydrogenase (NAD+) activity; IDA:UniProtKB. DR GO; GO:0004488; F:methylenetetrahydrofolate dehydrogenase (NADP+) activity; IDA:UniProtKB. DR GO; GO:0042301; F:phosphate ion binding; ISS:UniProtKB. DR GO; GO:0015943; P:formate biosynthetic process; IMP:MGI. DR GO; GO:0035999; P:tetrahydrofolate interconversion; IMP:MGI. DR GO; GO:0046653; P:tetrahydrofolate metabolic process; ISO:GO_Central. DR CDD; cd01080; NAD_bind_m-THF_DH_Cyclohyd; 1. DR FunFam; 3.40.50.10860:FF:000001; Bifunctional protein FolD; 1. DR FunFam; 3.40.50.720:FF:000070; probable bifunctional methylenetetrahydrofolate dehydrogenase/cyclohydrolase 2; 1. DR Gene3D; 3.40.50.10860; Leucine Dehydrogenase, chain A, domain 1; 1. DR Gene3D; 3.40.50.720; NAD(P)-binding Rossmann-like Domain; 1. DR HAMAP; MF_01576; THF_DHG_CYH; 1. DR InterPro; IPR046346; Aminoacid_DH-like_N_sf. DR InterPro; IPR036291; NAD(P)-bd_dom_sf. DR InterPro; IPR000672; THF_DH/CycHdrlase. DR InterPro; IPR020630; THF_DH/CycHdrlase_cat_dom. DR InterPro; IPR020867; THF_DH/CycHdrlase_CS. DR InterPro; IPR020631; THF_DH/CycHdrlase_NAD-bd_dom. DR PANTHER; PTHR48099:SF15; BIFUNCTIONAL METHYLENETETRAHYDROFOLATE DEHYDROGENASE_CYCLOHYDROLASE, MITOCHONDRIAL; 1. DR PANTHER; PTHR48099; C-1-TETRAHYDROFOLATE SYNTHASE, CYTOPLASMIC-RELATED; 1. DR Pfam; PF00763; THF_DHG_CYH; 1. DR Pfam; PF02882; THF_DHG_CYH_C; 1. DR PRINTS; PR00085; THFDHDRGNASE. DR SUPFAM; SSF53223; Aminoacid dehydrogenase-like, N-terminal domain; 1. DR SUPFAM; SSF51735; NAD(P)-binding Rossmann-fold domains; 1. DR PROSITE; PS00766; THF_DHG_CYH_1; 1. DR PROSITE; PS00767; THF_DHG_CYH_2; 1. PE 1: Evidence at protein level; KW Acetylation; Direct protein sequencing; Hydrolase; Isopeptide bond; KW Magnesium; Mitochondrion; Multifunctional enzyme; NAD; NADP; KW One-carbon metabolism; Oxidoreductase; Reference proteome; Transit peptide; KW Ubl conjugation. FT TRANSIT 1..35 FT /note="Mitochondrion" FT /evidence="ECO:0000269|PubMed:2647744" FT CHAIN 36..350 FT /note="Bifunctional methylenetetrahydrofolate FT dehydrogenase/cyclohydrolase, mitochondrial" FT /id="PRO_0000034050" FT BINDING 84..88 FT /ligand="substrate" FT /evidence="ECO:0000250|UniProtKB:P13995" FT BINDING 131..133 FT /ligand="substrate" FT /evidence="ECO:0000250|UniProtKB:P13995" FT BINDING 200..202 FT /ligand="NAD(+)" FT /ligand_id="ChEBI:CHEBI:57540" FT /evidence="ECO:0000250|UniProtKB:P13995" FT BINDING 233 FT /ligand="NAD(+)" FT /ligand_id="ChEBI:CHEBI:57540" FT /evidence="ECO:0000250|UniProtKB:P13995" FT BINDING 309..313 FT /ligand="substrate" FT /evidence="ECO:0000250|UniProtKB:P13995" FT MOD_RES 50 FT /note="N6-acetyllysine; alternate" FT /evidence="ECO:0000250|UniProtKB:P13995" FT CROSSLNK 50 FT /note="Glycyl lysine isopeptide (Lys-Gly) (interchain with FT G-Cter in SUMO2); alternate" FT /evidence="ECO:0000250|UniProtKB:P13995" SQ SEQUENCE 350 AA; 37863 MW; 896AD40D9154E9D7 CRC64; MASVSLLSAL AVRLLRPTHG CHPRLQPFHL AAVRNEAVVI SGRKLAQQIK QEVQQEVEEW VASGNKRPHL SVILVGDNPA SHSYVLNKTR AAAEVGINSE TIVKPASVSE EELLNSIRKL NNDENVDGLL VQLPLPEHID ERKVCNAVSP DKDVDGFHVI NVGRMCLDQY SMLPATPWGV WEIIKRTGIP TLGKNVVVAG RSKNVGMPIA MLLHTDGAHE RPGGDATVTI SHRYTPKEQL KKHTILADIV ISAAGIPNLI TADMIKEGAA VIDVGINRVQ DPVTAKPKLV GDVDFEGVKK KAGYITPVPG GVGPMTVAML MKNTIIAAKK VLRPEELEVF KSKQRGVATN //