ID HADB_CLODI Reviewed; 408 AA. AC Q5U924; DT 16-MAR-2016, integrated into UniProtKB/Swiss-Prot. DT 07-DEC-2004, sequence version 1. DT 18-JAN-2017, entry version 49. DE RecName: Full=(R)-2-hydroxyisocaproyl-CoA dehydratase alpha subunit {ECO:0000303|PubMed:15654892}; DE EC=4.2.1.157 {ECO:0000269|PubMed:15654892}; GN Name=hadB {ECO:0000303|PubMed:15654892}; GN ORFNames=CD630DERM_03970, ERS445050_03310; OS Clostridioides difficile (Peptoclostridium difficile). OG Plasmid 2. OC Bacteria; Firmicutes; Clostridia; Clostridiales; OC Peptostreptococcaceae; Clostridioides. OX NCBI_TaxID=1496; RN [1] RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], PROTEIN SEQUENCE OF 2-11, FUNCTION, RP CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, ENZYME REGULATION, RP COFACTOR, SUBUNIT, AND REACTION MECHANISM. RX PubMed=15654892; DOI=10.1111/j.1742-4658.2004.04498.x; RA Kim J., Darley D., Buckel W.; RT "2-hydroxyisocaproyl-CoA dehydratase and its activator from RT Clostridium difficile."; RL FEBS J. 272:550-561(2005). RN [2] RP NUCLEOTIDE SEQUENCE [GENOMIC DNA]. RC STRAIN=630Derm; RA Magalhaes I.L.F., Oliveira U., Santos F.R., Vidigal T.H.D.A., RA Brescovit A.D., Santos A.J.; RL Submitted (SEP-2014) to the EMBL/GenBank/DDBJ databases. RN [3] RP X-RAY CRYSTALLOGRAPHY (1.82 ANGSTROMS) IN COMPLEX WITH IRON-SULFUR RP (4FE-4S) AND SUBSTRATE ANALOG, FUNCTION, COFACTOR, SUBUNIT, AND RP REACTION MECHANISM. RC PLASMID=2; RX PubMed=21366233; DOI=10.1021/ja1076537; RA Knauer S.H., Buckel W., Dobbek H.; RT "Structural basis for reductive radical formation and electron RT recycling in (R)-2-hydroxyisocaproyl-CoA dehydratase."; RL J. Am. Chem. Soc. 133:4342-4347(2011). CC -!- FUNCTION: Involved in the reductive branch of L-leucine CC fermentation. Catalyzes the irreversible beta/alpha-elimination of CC water from (R)-2-hydroxyisocaproyl-CoA to yield isocaprenoyl-CoA. CC This beta/alpha-dehydration depends on the reductive formation of CC ketyl radicals on the substrate generated by injection of a single CC electron from the ATP-dependent activator protein HadI. The enzyme CC is specific for the R-isomer. {ECO:0000269|PubMed:15654892, CC ECO:0000269|PubMed:21366233}. CC -!- CATALYTIC ACTIVITY: (R)-2-hydroxy-4-methylpentanoyl-CoA = 4- CC methylpent-2-enoyl-CoA + H(2)O. {ECO:0000269|PubMed:15654892}. CC -!- COFACTOR: CC Name=[4Fe-4S] cluster; Xref=ChEBI:CHEBI:49883; CC Evidence={ECO:0000269|PubMed:15654892, CC ECO:0000269|PubMed:21366233}; CC Note=Binds 1 [4Fe-4S] cluster. {ECO:0000269|PubMed:15654892, CC ECO:0000269|PubMed:21366233}; CC -!- ENZYME REGULATION: Activated by HadI. CC {ECO:0000269|PubMed:15654892}. CC -!- BIOPHYSICOCHEMICAL PROPERTIES: CC Kinetic parameters: CC KM=0.05 mM for (R)-2-hydroxyisocaproyl-CoA CC {ECO:0000269|PubMed:15654892}; CC -!- SUBUNIT: Part of the heterodimeric complex HadBC composed of (R)- CC 2-hydroxyisocaproyl-CoA dehydratase alpha (HadB) and beta (HadC) CC subunit. {ECO:0000269|PubMed:15654892, CC ECO:0000269|PubMed:21366233}. CC -!- SIMILARITY: Belongs to the FldB/FldC dehydratase alpha/beta CC subunit 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; AY772816; AAV40819.1; -; Genomic_DNA. DR EMBL; LN614756; CEJ96915.1; -; Genomic_DNA. DR EMBL; LN831031; CKH72398.1; -; Genomic_DNA. DR PDB; 3O3M; X-ray; 1.82 A; A/C=1-408. DR PDB; 3O3N; X-ray; 2.30 A; A/C=1-408. DR PDB; 3O3O; X-ray; 2.00 A; A=1-408. DR PDBsum; 3O3M; -. DR PDBsum; 3O3N; -. DR PDBsum; 3O3O; -. DR SMR; Q5U924; -. DR STRING; 272563.CD0397; -. DR eggNOG; ENOG4105CMH; Bacteria. DR eggNOG; COG1775; LUCA. DR GO; GO:0051539; F:4 iron, 4 sulfur cluster binding; IDA:UniProtKB. DR GO; GO:0016836; F:hydro-lyase activity; IDA:UniProtKB. DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW. DR GO; GO:0006551; P:leucine metabolic process; IDA:UniProtKB. DR InterPro; IPR010327; FldB/FldC_alpha/beta. DR Pfam; PF06050; HGD-D; 1. PE 1: Evidence at protein level; KW 3D-structure; 4Fe-4S; Direct protein sequencing; Iron; Iron-sulfur; KW Lyase; Metal-binding; Plasmid. FT INIT_MET 1 1 Removed. {ECO:0000269|PubMed:15654892}. FT CHAIN 2 408 (R)-2-hydroxyisocaproyl-CoA dehydratase FT alpha subunit. FT /FTId=PRO_0000435662. FT METAL 84 84 Iron-sulfur (4Fe-4S). FT {ECO:0000269|PubMed:21366233}. FT METAL 117 117 Iron-sulfur (4Fe-4S). FT {ECO:0000269|PubMed:21366233}. FT METAL 346 346 Iron-sulfur (4Fe-4S). FT {ECO:0000269|PubMed:21366233}. FT BINDING 55 55 Substrate. {ECO:0000269|PubMed:21366233}. FT HELIX 7 28 {ECO:0000244|PDB:3O3M}. FT STRAND 32 35 {ECO:0000244|PDB:3O3M}. FT HELIX 42 45 {ECO:0000244|PDB:3O3M}. FT TURN 46 48 {ECO:0000244|PDB:3O3M}. FT STRAND 50 52 {ECO:0000244|PDB:3O3M}. FT HELIX 54 63 {ECO:0000244|PDB:3O3M}. FT HELIX 67 76 {ECO:0000244|PDB:3O3M}. FT HELIX 85 96 {ECO:0000244|PDB:3O3M}. FT STRAND 100 102 {ECO:0000244|PDB:3O3M}. FT STRAND 108 113 {ECO:0000244|PDB:3O3M}. FT HELIX 119 131 {ECO:0000244|PDB:3O3M}. FT STRAND 135 138 {ECO:0000244|PDB:3O3M}. FT STRAND 145 147 {ECO:0000244|PDB:3O3M}. FT HELIX 150 171 {ECO:0000244|PDB:3O3M}. FT HELIX 177 197 {ECO:0000244|PDB:3O3M}. FT TURN 198 203 {ECO:0000244|PDB:3O3M}. FT HELIX 211 213 {ECO:0000244|PDB:3O3M}. FT HELIX 214 223 {ECO:0000244|PDB:3O3M}. FT HELIX 228 246 {ECO:0000244|PDB:3O3M}. FT STRAND 258 264 {ECO:0000244|PDB:3O3M}. FT HELIX 268 270 {ECO:0000244|PDB:3O3M}. FT HELIX 271 280 {ECO:0000244|PDB:3O3M}. FT STRAND 283 287 {ECO:0000244|PDB:3O3M}. FT HELIX 290 292 {ECO:0000244|PDB:3O3M}. FT TURN 293 295 {ECO:0000244|PDB:3O3M}. FT HELIX 303 311 {ECO:0000244|PDB:3O3M}. FT HELIX 314 316 {ECO:0000244|PDB:3O3M}. FT HELIX 319 332 {ECO:0000244|PDB:3O3M}. FT STRAND 336 345 {ECO:0000244|PDB:3O3M}. FT HELIX 347 350 {ECO:0000244|PDB:3O3M}. FT HELIX 353 364 {ECO:0000244|PDB:3O3M}. FT STRAND 368 373 {ECO:0000244|PDB:3O3M}. FT HELIX 378 380 {ECO:0000244|PDB:3O3M}. FT HELIX 383 401 {ECO:0000244|PDB:3O3M}. SQ SEQUENCE 408 AA; 46334 MW; 702A372CB6796AB5 CRC64; MSEKKEARVV INDLLAEQYA NAFKAKEEGR PVGWSTSVFP QELAEVFDLN VLYPENQAAG VAAKKGSLEL CEIAESKGYS IDLCAYARTN FGLLENGGCE ALDMPAPDFL LCCNNICNQV IKWYENISRE LDIPLIMIDT TFNNEDEVTQ SRIDYIKAQF EEAIKQLEII SGKKFDPKKF EEVMKISAEN GRLWKYSMSL PADSSPSPMN GFDLFTYMAV IVCARGKKET TEAFKLLIEE LEDNMKTGKS SFRGEEKYRI MMEGIPCWPY IGYKMKTLAK FGVNMTGSVY PHAWALQYEV NDLDGMAVAY STMFNNVNLD RMTKYRVDSL VEGKCDGAFY HMNRSCKLMS LIQYEMQRRA AEETGLPYAG FDGDQADPRA FTNAQFETRI QGLVEVMEER KKLNRGEI //