ID ISPH_ECOLI Reviewed; 316 AA. AC P62623; P22565; DT 19-JUL-2004, integrated into UniProtKB/Swiss-Prot. DT 19-JUL-2004, sequence version 1. DT 27-MAR-2024, entry version 150. DE RecName: Full=4-hydroxy-3-methylbut-2-enyl diphosphate reductase {ECO:0000303|PubMed:12706830}; DE Short=HMBPP reductase {ECO:0000303|PubMed:12706830}; DE EC=1.17.7.4 {ECO:0000269|PubMed:12198182, ECO:0000269|PubMed:12571359, ECO:0000269|PubMed:12706830}; DE AltName: Full=1-hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate reductase {ECO:0000303|PubMed:12198182}; GN Name=ispH {ECO:0000303|PubMed:11818558}; Synonyms=lytB, yaaE; GN OrderedLocusNames=b0029, JW0027; OS Escherichia coli (strain K12). OC Bacteria; Pseudomonadota; Gammaproteobacteria; Enterobacterales; OC Enterobacteriaceae; Escherichia. OX NCBI_TaxID=83333; RN [1] RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], FUNCTION, AND PATHWAY. RX PubMed=11818558; DOI=10.1073/pnas.032658999; RA Rohdich F., Hecht S., Gaertner K., Adam P., Krieger C., Amslinger S., RA Arigoni D., Bacher A., Eisenreich W.; RT "Studies on the nonmevalonate terpene biosynthetic pathway: metabolic role RT of IspH (LytB) protein."; RL Proc. Natl. Acad. Sci. U.S.A. 99:1158-1163(2002). RN [2] RP NUCLEOTIDE SEQUENCE [GENOMIC DNA]. RC STRAIN=K12; RX PubMed=2011499; DOI=10.1093/nar/19.1.180; RA Bouvier J., Stragier P.; RT "Nucleotide sequence of the lsp-dapB interval in Escherichia coli."; RL Nucleic Acids Res. 19:180-180(1991). RN [3] RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA]. RC STRAIN=K12; RX PubMed=1630901; DOI=10.1093/nar/20.13.3305; RA Yura T., Mori H., Nagai H., Nagata T., Ishihama A., Fujita N., Isono K., RA Mizobuchi K., Nakata A.; RT "Systematic sequencing of the Escherichia coli genome: analysis of the RT 0-2.4 min region."; RL Nucleic Acids Res. 20:3305-3308(1992). RN [4] 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 [5] 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 [6] RP IDENTIFICATION. RX PubMed=8432714; DOI=10.1128/jb.175.4.1203-1205.1993; RA Gustafson C.E., Kaul S., Ishiguro E.E.; RT "Identification of the Escherichia coli lytB gene, which is involved in RT penicillin tolerance and control of the stringent response."; RL J. Bacteriol. 175:1203-1205(1993). RN [7] RP FUNCTION, PATHWAY, AND DISRUPTION PHENOTYPE. RC STRAIN=K12 / ATCC 23716 / DSM 498 / CIP 110067 / IMG 1711; RX PubMed=11418107; DOI=10.1016/s0014-5793(01)02516-9; RA Altincicek B., Kollas A.-K., Eberl M., Wiesner J., Sanderbrand S., RA Hintz M., Beck E., Jomaa H.; RT "LytB, a novel gene of the 2-C-methyl-D-erythritol 4-phosphate pathway of RT isoprenoid biosynthesis in Escherichia coli."; RL FEBS Lett. 499:37-40(2001). RN [8] RP DISRUPTION PHENOTYPE. RX PubMed=11717301; DOI=10.1128/jb.183.24.7403-7407.2001; RA McAteer S., Coulson A., McLennan N., Masters M.; RT "The lytB gene of Escherichia coli is essential and specifies a product RT needed for isoprenoid biosynthesis."; RL J. Bacteriol. 183:7403-7407(2001). RN [9] RP CATALYTIC ACTIVITY. RX PubMed=12198182; DOI=10.1073/pnas.182412599; RA Adam P., Hecht S., Eisenreich W., Kaiser J., Graewert T., Arigoni D., RA Bacher A., Rohdich F.; RT "Biosynthesis of terpenes: studies on 1-hydroxy-2-methyl-2-(E)-butenyl 4- RT diphosphate reductase."; RL Proc. Natl. Acad. Sci. U.S.A. 99:12108-12113(2002). RN [10] RP FUNCTION, CATALYTIC ACTIVITY, COFACTOR, AND SUBUNIT. RX PubMed=12706830; DOI=10.1016/s0014-5793(03)00317-x; RA Wolff M., Seemann M., Tse Sum Bui B., Frapart Y., Tritsch D., RA Garcia Estrabot A., Rodriguez-Concepcion M., Boronat A., Marquet A., RA Rohmer M.; RT "Isoprenoid biosynthesis via the methylerythritol phosphate pathway: the RT (E)-4-hydroxy-3-methylbut-2-enyl diphosphate reductase (LytB/IspH) from RT Escherichia coli is a [4Fe-4S] protein."; RL FEBS Lett. 541:115-120(2003). RN [11] RP CATALYTIC ACTIVITY. RX PubMed=12571359; DOI=10.1073/pnas.0337742100; RA Rohdich F., Zepeck F., Adam P., Hecht S., Kaiser J., Laupitz R., RA Graewert T., Amslinger S., Eisenreich W., Bacher A., Arigoni D.; RT "The deoxyxylulose phosphate pathway of isoprenoid biosynthesis: studies on RT the mechanisms of the reactions catalyzed by IspG and IspH protein."; RL Proc. Natl. Acad. Sci. U.S.A. 100:1586-1591(2003). RN [12] RP PRESENCE OF A 3FE-4S IRON-SULFUR CLUSTER, ACTIVITY REGULATION, KINETIC RP PARAMETERS, AND MUTAGENESIS OF CYS-12; CYS-96 AND CYS-197. RX PubMed=15469281; DOI=10.1021/ja0471727; RA Graewert T., Kaiser J., Zepeck F., Laupitz R., Hecht S., Amslinger S., RA Schramek N., Schleicher E., Weber S., Haslbeck M., Buchner J., Rieder C., RA Arigoni D., Bacher A., Eisenreich W., Rohdich F.; RT "IspH protein of Escherichia coli: studies on iron-sulfur cluster RT implementation and catalysis."; RL J. Am. Chem. Soc. 126:12847-12855(2004). RN [13] RP ACTIVITY REGULATION, AND COFACTOR. RX PubMed=22012762; DOI=10.1002/anie.201104562; RA Ahrens-Botzong A., Janthawornpong K., Wolny J.A., Tambou E.N., Rohmer M., RA Krasutsky S., Poulter C.D., Schuenemann V., Seemann M.; RT "Biosynthesis of isoprene units: Moessbauer spectroscopy of substrate and RT inhibitor binding to the [4Fe-4S] cluster of the LytB/IspH enzyme."; RL Angew. Chem. Int. Ed. 50:11976-11979(2011). RN [14] {ECO:0007744|PDB:3F7T} RP X-RAY CRYSTALLOGRAPHY (1.80 ANGSTROMS) IN COMPLEX WITH IRON-SULFUR RP (3FE-4S), FUNCTION, COFACTOR, AND MUTAGENESIS OF HIS-41; HIS-74; VAL-99; RP HIS-124; GLU-126; THR-167; SER-225 AND ASN-227. RX PubMed=19569147; DOI=10.1002/anie.200900548; RA Graewert T., Rohdich F., Span I., Bacher A., Eisenreich W., Eppinger J., RA Groll M.; RT "Structure of active IspH enzyme from Escherichia coli provides mechanistic RT insights into substrate reduction."; RL Angew. Chem. Int. Ed. Engl. 48:5756-5759(2009). RN [15] {ECO:0007744|PDB:3KE8, ECO:0007744|PDB:3KE9, ECO:0007744|PDB:3KEF, ECO:0007744|PDB:3KEL, ECO:0007744|PDB:3KEM} RP X-RAY CRYSTALLOGRAPHY (1.70 ANGSTROMS) IN COMPLEXES WITH SUBSTRATE; RP PRODUCTS; IRON-SULFUR (3FE-4S) AND IRON-SULFUR (4FE-4S), COFACTOR, AND RP MUTAGENESIS OF CYS-12; CYS-96 AND CYS-197. RC STRAIN=K12; RX PubMed=20080550; DOI=10.1073/pnas.0913045107; RA Graewert T., Span I., Eisenreich W., Rohdich F., Eppinger J., Bacher A., RA Groll M.; RT "Probing the reaction mechanism of IspH protein by x-ray structure RT analysis."; RL Proc. Natl. Acad. Sci. U.S.A. 107:1077-1081(2010). RN [16] {ECO:0007744|PDB:4EB3} RP X-RAY CRYSTALLOGRAPHY (1.90 ANGSTROMS) OF 1-315 IN COMPLEX WITH ISO-HMBPP RP AND IRON-SULFUR (4FE-4S), COFACTOR, REACTION MECHANISM, AND ACTIVE SITE. RX PubMed=22687151; DOI=10.1021/ja303445z; RA Wang W., Wang K., Span I., Jauch J., Bacher A., Groll M., Oldfield E.; RT "Are free radicals involved in IspH catalysis? An EPR and crystallographic RT investigation."; RL J. Am. Chem. Soc. 134:11225-11234(2012). RN [17] {ECO:0007744|PDB:3SZL, ECO:0007744|PDB:3SZO, ECO:0007744|PDB:3SZU, ECO:0007744|PDB:3T0F, ECO:0007744|PDB:3T0G} RP X-RAY CRYSTALLOGRAPHY (1.40 ANGSTROMS) OF WILD-TYPE AND MUTANTS CYS-167; RP ASP-126 AND GLN-126 IN COMPLEXES WITH SUBSTRATE AND IRON-SULFUR (4FE-4S), RP FUNCTION, COFACTOR, MUTAGENESIS OF GLU-126 AND THR-167, REACTION MECHANISM, RP AND ACTIVE SITE. RX PubMed=22137895; DOI=10.1016/j.jmb.2011.11.033; RA Span I., Graewert T., Bacher A., Eisenreich W., Groll M.; RT "Crystal structures of mutant IspH proteins reveal a rotation of the RT substrate's hydroxymethyl group during catalysis."; RL J. Mol. Biol. 416:1-9(2012). RN [18] RP X-RAY CRYSTALLOGRAPHY (1.50 ANGSTROMS) IN COMPLEXES WITH VARIOUS COMPOUNDS; RP IRON-SULFUR (3FE-4S) AND IRON-SULFUR (4FE-4S), FUNCTION, AND ACETYLENE RP HYDRATASE ACTIVITY. RX PubMed=22948824; DOI=10.1038/ncomms2052; RA Span I., Wang K., Wang W., Zhang Y., Bacher A., Eisenreich W., Li K., RA Schulz C., Oldfield E., Groll M.; RT "Discovery of acetylene hydratase activity of the iron-sulphur protein RT IspH."; RL Nat. Commun. 3:1042-1042(2012). RN [19] RP X-RAY CRYSTALLOGRAPHY (1.68 ANGSTROMS) IN COMPLEX WITH SUBSTRATE ANALOGS RP AND IRON-SULFUR (4FE-4S). RA Borel F., Barbier E., Kratsutsky S., Janthawornpong K., Rohmer M., RA Dale Poulter C., Ferrer J.L., Seemann M.; RT "Crystal structures of Escherichia coli Isph in complex with two potent RT inhibitors of the methylerythritol phosphate pathway, a target for the RT development of new antibacterial and antiparasitic drugs."; RL Submitted (DEC-2012) to the PDB data bank. RN [20] RP X-RAY CRYSTALLOGRAPHY (1.35 ANGSTROMS) OF 1-315 IN COMPLEXES WITH SUBSTRATE RP ANALOGS AND IRON-SULFUR (4FE-4S). RX PubMed=23307751; DOI=10.1002/anie.201208469; RA Span I., Wang K., Wang W., Jauch J., Eisenreich W., Bacher A., Oldfield E., RA Groll M.; RT "Structures of fluoro, amino, and thiol inhibitors bound to the [Fe4S4] RT protein IspH."; RL Angew. Chem. Int. Ed. Engl. 52:2118-2121(2013). CC -!- FUNCTION: Catalyzes the conversion of 1-hydroxy-2-methyl-2-(E)-butenyl CC 4-diphosphate (HMBPP) into a mixture of isopentenyl diphosphate (IPP) CC and dimethylallyl diphosphate (DMAPP). Acts in the terminal step of the CC DOXP/MEP pathway for isoprenoid precursor biosynthesis CC (PubMed:11818558, PubMed:11418107, PubMed:12706830, PubMed:19569147, CC PubMed:22137895). In vitro, can also hydrate acetylenes to aldehydes CC and ketones via anti-Markovnikov/Markovnikov addition CC (PubMed:22948824). {ECO:0000269|PubMed:11418107, CC ECO:0000269|PubMed:11818558, ECO:0000269|PubMed:12706830, CC ECO:0000269|PubMed:19569147, ECO:0000269|PubMed:22137895, CC ECO:0000269|PubMed:22948824}. CC -!- CATALYTIC ACTIVITY: CC Reaction=H2O + isopentenyl diphosphate + 2 oxidized [2Fe-2S]- CC [ferredoxin] = (2E)-4-hydroxy-3-methylbut-2-enyl diphosphate + 2 H(+) CC + 2 reduced [2Fe-2S]-[ferredoxin]; Xref=Rhea:RHEA:24488, Rhea:RHEA- CC COMP:10000, Rhea:RHEA-COMP:10001, ChEBI:CHEBI:15377, CC ChEBI:CHEBI:15378, ChEBI:CHEBI:33737, ChEBI:CHEBI:33738, CC ChEBI:CHEBI:128753, ChEBI:CHEBI:128769; EC=1.17.7.4; CC Evidence={ECO:0000269|PubMed:12198182, ECO:0000269|PubMed:12571359, CC ECO:0000269|PubMed:12706830}; CC -!- CATALYTIC ACTIVITY: CC Reaction=dimethylallyl diphosphate + H2O + 2 oxidized [2Fe-2S]- CC [ferredoxin] = (2E)-4-hydroxy-3-methylbut-2-enyl diphosphate + 2 H(+) CC + 2 reduced [2Fe-2S]-[ferredoxin]; Xref=Rhea:RHEA:24825, Rhea:RHEA- CC COMP:10000, Rhea:RHEA-COMP:10001, ChEBI:CHEBI:15377, CC ChEBI:CHEBI:15378, ChEBI:CHEBI:33737, ChEBI:CHEBI:33738, CC ChEBI:CHEBI:57623, ChEBI:CHEBI:128753; EC=1.17.7.4; CC Evidence={ECO:0000269|PubMed:12198182, ECO:0000269|PubMed:12571359, CC ECO:0000269|PubMed:12706830}; CC -!- COFACTOR: CC Name=[4Fe-4S] cluster; Xref=ChEBI:CHEBI:49883; CC Evidence={ECO:0000269|PubMed:12706830, ECO:0000269|PubMed:20080550, CC ECO:0000269|PubMed:22012762, ECO:0000269|PubMed:22137895, CC ECO:0000269|PubMed:22687151}; CC Note=Was shown to bind 1 [3Fe-4S] cluster per subunit (PubMed:19569147, CC PubMed:20080550, PubMed:15469281). However, it initially contains a CC [4Fe-4S] cluster which easily degrades into a [3Fe-4S] form in the CC presence of oxygen (PubMed:12706830, PubMed:22137895, PubMed:20080550, CC PubMed:22687151). {ECO:0000269|PubMed:12706830, CC ECO:0000269|PubMed:15469281, ECO:0000269|PubMed:19569147, CC ECO:0000269|PubMed:20080550, ECO:0000269|PubMed:22137895, CC ECO:0000269|PubMed:22687151}; CC -!- ACTIVITY REGULATION: Addition of Ca(2+), Mg(2+), Mn(2+), Ni(2+), Co(2+) CC or Fe(2+) decreases the catalytic activity (PubMed:15469281). Addition CC of Zn(2+) results in complete loss of activity (PubMed:15469281). Is CC potently inhibited by substrate analogs in which the hydroxy group in CC HMBPP is replaced by an amino or thiol group (PubMed:22012762). CC {ECO:0000269|PubMed:15469281, ECO:0000269|PubMed:22012762}. CC -!- BIOPHYSICOCHEMICAL PROPERTIES: CC Kinetic parameters: CC KM=30 uM for 1-hydroxy-2-methyl-2-(E)-butenyl 4-diphosphate CC {ECO:0000269|PubMed:15469281}; CC pH dependence: CC Optimum pH is 7.0.; CC -!- PATHWAY: Isoprenoid biosynthesis; dimethylallyl diphosphate CC biosynthesis; dimethylallyl diphosphate from (2E)-4-hydroxy-3- CC methylbutenyl diphosphate: step 1/1. {ECO:0000269|PubMed:11818558, CC ECO:0000305|PubMed:11418107}. CC -!- PATHWAY: Isoprenoid biosynthesis; isopentenyl diphosphate biosynthesis CC via DXP pathway; isopentenyl diphosphate from 1-deoxy-D-xylulose 5- CC phosphate: step 6/6. {ECO:0000269|PubMed:11818558, CC ECO:0000305|PubMed:11418107}. CC -!- SUBUNIT: Homodimer. {ECO:0000269|PubMed:12706830}. CC -!- DISRUPTION PHENOTYPE: Cells lacking this gene are viable only if the CC medium is supplemented with mevalonate or the cells are complemented CC with an episomal copy of ispH (PubMed:11418107). A conditional E.coli CC lytB mutant shows that LytB is essential for survival and that CC depletion of LytB results in cell lysis (PubMed:11717301). CC {ECO:0000269|PubMed:11418107, ECO:0000269|PubMed:11717301}. CC -!- SIMILARITY: Belongs to the IspH family. {ECO:0000255|HAMAP- CC Rule:MF_00191}. 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; AY062212; AAL38655.1; -; Genomic_DNA. DR EMBL; X54945; CAA38707.1; -; Genomic_DNA. DR EMBL; U00096; AAC73140.1; -; Genomic_DNA. DR EMBL; AP009048; BAB96598.1; -; Genomic_DNA. DR PIR; JE0403; JE0403. DR RefSeq; NP_414570.1; NC_000913.3. DR RefSeq; WP_001166395.1; NZ_SSZK01000004.1. DR PDB; 3F7T; X-ray; 1.80 A; A/B=1-316. DR PDB; 3KE8; X-ray; 1.70 A; A/B=1-316. DR PDB; 3KE9; X-ray; 1.90 A; A/B=1-316. DR PDB; 3KEF; X-ray; 1.70 A; A/B=1-316. DR PDB; 3KEL; X-ray; 1.80 A; A/B=1-316. DR PDB; 3KEM; X-ray; 2.00 A; A/B=1-316. DR PDB; 3SZL; X-ray; 1.60 A; A/B=1-316. DR PDB; 3SZO; X-ray; 1.60 A; A/B=1-316. DR PDB; 3SZU; X-ray; 1.40 A; A/B=1-316. DR PDB; 3T0F; X-ray; 1.90 A; A/B=1-316. DR PDB; 3T0G; X-ray; 2.10 A; A/B=1-316. DR PDB; 3URK; X-ray; 1.50 A; A/B=1-316. DR PDB; 3UTC; X-ray; 1.90 A; A/B=1-316. DR PDB; 3UTD; X-ray; 1.70 A; A/B=1-316. DR PDB; 3UV3; X-ray; 1.60 A; A/B=1-316. DR PDB; 3UV6; X-ray; 1.70 A; A/B=1-316. DR PDB; 3UV7; X-ray; 1.60 A; A/B=1-316. DR PDB; 3UWM; X-ray; 1.80 A; A/B=1-316. DR PDB; 3ZGL; X-ray; 1.68 A; A/B=1-316. DR PDB; 3ZGN; X-ray; 1.95 A; A/B=1-316. DR PDB; 4EB3; X-ray; 1.90 A; A/B=1-315. DR PDB; 4H4C; X-ray; 1.80 A; A/B=1-315. DR PDB; 4H4D; X-ray; 1.35 A; A/B=1-315. DR PDB; 4H4E; X-ray; 1.70 A; A/B=1-315. DR PDBsum; 3F7T; -. DR PDBsum; 3KE8; -. DR PDBsum; 3KE9; -. DR PDBsum; 3KEF; -. DR PDBsum; 3KEL; -. DR PDBsum; 3KEM; -. DR PDBsum; 3SZL; -. DR PDBsum; 3SZO; -. DR PDBsum; 3SZU; -. DR PDBsum; 3T0F; -. DR PDBsum; 3T0G; -. DR PDBsum; 3URK; -. DR PDBsum; 3UTC; -. DR PDBsum; 3UTD; -. DR PDBsum; 3UV3; -. DR PDBsum; 3UV6; -. DR PDBsum; 3UV7; -. DR PDBsum; 3UWM; -. DR PDBsum; 3ZGL; -. DR PDBsum; 3ZGN; -. DR PDBsum; 4EB3; -. DR PDBsum; 4H4C; -. DR PDBsum; 4H4D; -. DR PDBsum; 4H4E; -. DR AlphaFoldDB; P62623; -. DR SMR; P62623; -. DR BioGRID; 4262917; 331. DR BioGRID; 849179; 2. DR DIP; DIP-35808N; -. DR IntAct; P62623; 7. DR STRING; 511145.b0029; -. DR BindingDB; P62623; -. DR DrugBank; DB01785; Dimethylallyl Diphosphate. DR DrugBank; DB04714; ISOPENTENYL PYROPHOSPHATE. DR jPOST; P62623; -. DR PaxDb; 511145-b0029; -. DR EnsemblBacteria; AAC73140; AAC73140; b0029. DR GeneID; 75203956; -. DR GeneID; 944777; -. DR KEGG; ecj:JW0027; -. DR KEGG; eco:b0029; -. DR PATRIC; fig|1411691.4.peg.2256; -. DR EchoBASE; EB1073; -. DR eggNOG; COG0761; Bacteria. DR HOGENOM; CLU_027486_1_0_6; -. DR InParanoid; P62623; -. DR OMA; SEMIHNP; -. DR OrthoDB; 9804068at2; -. DR PhylomeDB; P62623; -. DR BioCyc; EcoCyc:EG11081-MONOMER; -. DR BioCyc; MetaCyc:EG11081-MONOMER; -. DR UniPathway; UPA00056; UER00097. DR UniPathway; UPA00059; UER00105. DR EvolutionaryTrace; P62623; -. DR PRO; PR:P62623; -. DR Proteomes; UP000000318; Chromosome. DR Proteomes; UP000000625; Chromosome. DR GO; GO:0005829; C:cytosol; IDA:EcoCyc. DR GO; GO:0051538; F:3 iron, 4 sulfur cluster binding; IDA:EcoCyc. DR GO; GO:0051539; F:4 iron, 4 sulfur cluster binding; IEA:UniProtKB-UniRule. DR GO; GO:0051745; F:4-hydroxy-3-methylbut-2-en-1-yl diphosphate reductase activity; IDA:EcoCyc. DR GO; GO:0042380; F:hydroxymethylbutenyl pyrophosphate reductase activity; IDA:EcoCyc. DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW. DR GO; GO:0050992; P:dimethylallyl diphosphate biosynthetic process; IEA:UniProtKB-UniRule. DR GO; GO:0019288; P:isopentenyl diphosphate biosynthetic process, methylerythritol 4-phosphate pathway; IMP:EcoCyc. DR GO; GO:0016114; P:terpenoid biosynthetic process; IEA:UniProtKB-UniRule. DR CDD; cd13944; lytB_ispH; 1. DR Gene3D; 3.40.50.11270; -; 1. DR Gene3D; 3.40.1010.20; 4-hydroxy-3-methylbut-2-enyl diphosphate reductase, catalytic domain; 2. DR HAMAP; MF_00191; IspH; 1. DR InterPro; IPR003451; LytB/IspH. DR NCBIfam; TIGR00216; ispH_lytB; 1. DR PANTHER; PTHR30426; 4-HYDROXY-3-METHYLBUT-2-ENYL DIPHOSPHATE REDUCTASE; 1. DR PANTHER; PTHR30426:SF0; 4-HYDROXY-3-METHYLBUT-2-ENYL DIPHOSPHATE REDUCTASE; 1. DR Pfam; PF02401; LYTB; 1. PE 1: Evidence at protein level; KW 3D-structure; 4Fe-4S; Iron; Iron-sulfur; Isoprene biosynthesis; KW Metal-binding; Oxidoreductase; Reference proteome. FT CHAIN 1..316 FT /note="4-hydroxy-3-methylbut-2-enyl diphosphate reductase" FT /id="PRO_0000128812" FT ACT_SITE 126 FT /note="Proton donor" FT /evidence="ECO:0000305|PubMed:22137895, FT ECO:0000305|PubMed:22687151" FT BINDING 12 FT /ligand="[4Fe-4S] cluster" FT /ligand_id="ChEBI:CHEBI:49883" FT /evidence="ECO:0000269|PubMed:20080550, FT ECO:0000269|PubMed:22137895, ECO:0000269|PubMed:22687151, FT ECO:0000269|PubMed:22948824, ECO:0000269|PubMed:23307751, FT ECO:0007744|PDB:3KE8, ECO:0007744|PDB:3SZL, FT ECO:0007744|PDB:3URK, ECO:0007744|PDB:4EB3, FT ECO:0007744|PDB:4H4C" FT BINDING 41 FT /ligand="substrate" FT /evidence="ECO:0000269|PubMed:20080550, FT ECO:0000269|PubMed:22137895, ECO:0000305|PubMed:22687151, FT ECO:0007744|PDB:3KE8, ECO:0007744|PDB:3SZL, FT ECO:0007744|PDB:4EB3" FT BINDING 74 FT /ligand="substrate" FT /evidence="ECO:0000269|PubMed:20080550, FT ECO:0000269|PubMed:22137895, ECO:0000305|PubMed:22687151, FT ECO:0007744|PDB:3KE8, ECO:0007744|PDB:3SZL, FT ECO:0007744|PDB:4EB3" FT BINDING 96 FT /ligand="[4Fe-4S] cluster" FT /ligand_id="ChEBI:CHEBI:49883" FT /evidence="ECO:0000269|PubMed:20080550, FT ECO:0000269|PubMed:22137895, ECO:0000269|PubMed:22687151, FT ECO:0000269|PubMed:22948824, ECO:0000269|PubMed:23307751, FT ECO:0007744|PDB:3KE8, ECO:0007744|PDB:3SZL, FT ECO:0007744|PDB:3URK, ECO:0007744|PDB:4EB3, FT ECO:0007744|PDB:4H4C" FT BINDING 124 FT /ligand="substrate" FT /evidence="ECO:0000269|PubMed:20080550, FT ECO:0000269|PubMed:22137895, ECO:0000305|PubMed:22687151, FT ECO:0007744|PDB:3KE8, ECO:0007744|PDB:3SZL, FT ECO:0007744|PDB:4EB3" FT BINDING 167 FT /ligand="substrate" FT /evidence="ECO:0000269|PubMed:20080550, FT ECO:0000269|PubMed:22137895, ECO:0000305|PubMed:22687151, FT ECO:0007744|PDB:3KE8, ECO:0007744|PDB:3SZL, FT ECO:0007744|PDB:4EB3" FT BINDING 197 FT /ligand="[4Fe-4S] cluster" FT /ligand_id="ChEBI:CHEBI:49883" FT /evidence="ECO:0000269|PubMed:20080550, FT ECO:0000269|PubMed:22137895, ECO:0000269|PubMed:22687151, FT ECO:0000269|PubMed:22948824, ECO:0000269|PubMed:23307751, FT ECO:0007744|PDB:3KE8, ECO:0007744|PDB:3SZL, FT ECO:0007744|PDB:3URK, ECO:0007744|PDB:4EB3, FT ECO:0007744|PDB:4H4C" FT BINDING 225..227 FT /ligand="substrate" FT /evidence="ECO:0000269|PubMed:20080550, FT ECO:0000269|PubMed:22137895, ECO:0000305|PubMed:22687151, FT ECO:0007744|PDB:3KE8, ECO:0007744|PDB:3SZL, FT ECO:0007744|PDB:4EB3" FT BINDING 269 FT /ligand="substrate" FT /evidence="ECO:0000269|PubMed:20080550, FT ECO:0000269|PubMed:22137895, ECO:0000305|PubMed:22687151, FT ECO:0007744|PDB:3KE8, ECO:0007744|PDB:3SZL, FT ECO:0007744|PDB:4EB3" FT MUTAGEN 12 FT /note="C->S: Loss of catalytic activity." FT /evidence="ECO:0000269|PubMed:15469281, FT ECO:0000269|PubMed:20080550" FT MUTAGEN 41 FT /note="H->N: No effect on catalytic activity." FT /evidence="ECO:0000269|PubMed:19569147" FT MUTAGEN 74 FT /note="H->N: Reduces catalytic activity 2-fold." FT /evidence="ECO:0000269|PubMed:19569147" FT MUTAGEN 96 FT /note="C->S: Loss of catalytic activity." FT /evidence="ECO:0000269|PubMed:15469281, FT ECO:0000269|PubMed:20080550" FT MUTAGEN 99 FT /note="V->A: No effect on catalytic activity." FT /evidence="ECO:0000269|PubMed:19569147" FT MUTAGEN 124 FT /note="H->N: Loss of catalytic activity." FT /evidence="ECO:0000269|PubMed:19569147" FT MUTAGEN 126 FT /note="E->D,Q: Loss of catalytic activity." FT /evidence="ECO:0000269|PubMed:19569147, FT ECO:0000269|PubMed:22137895" FT MUTAGEN 167 FT /note="T->C: Reduces catalytic activity 3-fold." FT /evidence="ECO:0000269|PubMed:19569147, FT ECO:0000269|PubMed:22137895" FT MUTAGEN 167 FT /note="T->S: No effect on catalytic activity." FT /evidence="ECO:0000269|PubMed:19569147, FT ECO:0000269|PubMed:22137895" FT MUTAGEN 197 FT /note="C->S: Loss of catalytic activity." FT /evidence="ECO:0000269|PubMed:15469281, FT ECO:0000269|PubMed:20080550" FT MUTAGEN 225 FT /note="S->C: Loss of catalytic activity." FT /evidence="ECO:0000269|PubMed:19569147" FT MUTAGEN 227 FT /note="N->Q: Reduces catalytic activity 20-fold." FT /evidence="ECO:0000269|PubMed:19569147" FT STRAND 2..5 FT /evidence="ECO:0007829|PDB:4H4D" FT HELIX 13..29 FT /evidence="ECO:0007829|PDB:4H4D" FT STRAND 33..37 FT /evidence="ECO:0007829|PDB:4H4D" FT STRAND 39..41 FT /evidence="ECO:0007829|PDB:4H4D" FT HELIX 43..51 FT /evidence="ECO:0007829|PDB:4H4D" FT STRAND 54..59 FT /evidence="ECO:0007829|PDB:4H4D" FT HELIX 60..62 FT /evidence="ECO:0007829|PDB:4H4D" FT STRAND 68..71 FT /evidence="ECO:0007829|PDB:4H4D" FT HELIX 78..86 FT /evidence="ECO:0007829|PDB:4H4D" FT STRAND 90..93 FT /evidence="ECO:0007829|PDB:4H4D" FT HELIX 97..112 FT /evidence="ECO:0007829|PDB:4H4D" FT STRAND 115..120 FT /evidence="ECO:0007829|PDB:4H4D" FT HELIX 125..131 FT /evidence="ECO:0007829|PDB:4H4D" FT STRAND 137..139 FT /evidence="ECO:0007829|PDB:3T0G" FT STRAND 141..144 FT /evidence="ECO:0007829|PDB:4H4D" FT HELIX 147..152 FT /evidence="ECO:0007829|PDB:4H4D" FT STRAND 160..165 FT /evidence="ECO:0007829|PDB:4H4D" FT STRAND 167..169 FT /evidence="ECO:0007829|PDB:3T0F" FT HELIX 171..184 FT /evidence="ECO:0007829|PDB:4H4D" FT STRAND 191..193 FT /evidence="ECO:0007829|PDB:3KEM" FT HELIX 198..213 FT /evidence="ECO:0007829|PDB:4H4D" FT STRAND 215..220 FT /evidence="ECO:0007829|PDB:4H4D" FT HELIX 226..237 FT /evidence="ECO:0007829|PDB:4H4D" FT STRAND 241..247 FT /evidence="ECO:0007829|PDB:4H4D" FT HELIX 248..250 FT /evidence="ECO:0007829|PDB:4H4D" FT HELIX 253..255 FT /evidence="ECO:0007829|PDB:4H4D" FT TURN 256..258 FT /evidence="ECO:0007829|PDB:4H4D" FT STRAND 260..266 FT /evidence="ECO:0007829|PDB:4H4D" FT HELIX 272..284 FT /evidence="ECO:0007829|PDB:4H4D" FT STRAND 289..292 FT /evidence="ECO:0007829|PDB:4H4D" FT STRAND 300..302 FT /evidence="ECO:0007829|PDB:4H4D" FT HELIX 306..308 FT /evidence="ECO:0007829|PDB:4H4D" SQ SEQUENCE 316 AA; 34775 MW; 0E7B378BD49AB771 CRC64; MQILLANPRG FCAGVDRAIS IVENALAIYG APIYVRHEVV HNRYVVDSLR ERGAIFIEQI SEVPDGAILI FSAHGVSQAV RNEAKSRDLT VFDATCPLVT KVHMEVARAS RRGEESILIG HAGHPEVEGT MGQYSNPEGG MYLVESPDDV WKLTVKNEEK LSFMTQTTLS VDDTSDVIDA LRKRFPKIVG PRKDDICYAT TNRQEAVRAL AEQAEVVLVV GSKNSSNSNR LAELAQRMGK RAFLIDDAKD IQEEWVKEVK CVGVTAGASA PDILVQNVVA RLQQLGGGEA IPLEGREENI VFEVPKELRV DIREVD //