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 03-MAY-2023, entry version 75. 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; Bacillota; Clostridia; Eubacteriales; Peptostreptococcaceae; OC Clostridioides. OX NCBI_TaxID=1496; RN [1] RP NUCLEOTIDE SEQUENCE [GENOMIC DNA], PROTEIN SEQUENCE OF 2-11, FUNCTION, RP CATALYTIC ACTIVITY, BIOPHYSICOCHEMICAL PROPERTIES, ACTIVITY 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 Clostridium RT 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 (4FE-4S) RP AND SUBSTRATE ANALOG, FUNCTION, COFACTOR, SUBUNIT, AND 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 recycling in RT (R)-2-hydroxyisocaproyl-CoA dehydratase."; RL J. Am. Chem. Soc. 133:4342-4347(2011). CC -!- FUNCTION: Involved in the reductive branch of L-leucine fermentation. CC Catalyzes the irreversible beta/alpha-elimination of water from (R)-2- CC hydroxyisocaproyl-CoA to yield isocaprenoyl-CoA. This beta/alpha- CC dehydration depends on the reductive formation of ketyl radicals on the CC substrate generated by injection of a single electron from the ATP- CC dependent activator protein HadI. The enzyme is specific for the R- CC isomer. {ECO:0000269|PubMed:15654892, ECO:0000269|PubMed:21366233}. CC -!- CATALYTIC ACTIVITY: CC Reaction=(R)-2-hydroxy-4-methylpentanoyl-CoA = 4-methylpent-2-enoyl-CoA CC + H2O; Xref=Rhea:RHEA:46924, ChEBI:CHEBI:15377, ChEBI:CHEBI:87119, CC ChEBI:CHEBI:87120; EC=4.2.1.157; CC Evidence={ECO:0000269|PubMed:15654892}; CC -!- COFACTOR: CC Name=[4Fe-4S] cluster; Xref=ChEBI:CHEBI:49883; CC Evidence={ECO:0000269|PubMed:15654892, ECO:0000269|PubMed:21366233}; CC Note=Binds 1 [4Fe-4S] cluster. {ECO:0000269|PubMed:15654892, CC ECO:0000269|PubMed:21366233}; CC -!- ACTIVITY REGULATION: Activated by HadI. {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)-2- CC hydroxyisocaproyl-CoA dehydratase alpha (HadB) and beta (HadC) subunit. CC {ECO:0000269|PubMed:15654892, ECO:0000269|PubMed:21366233}. CC -!- SIMILARITY: Belongs to the FldB/FldC dehydratase alpha/beta subunit 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; AY772816; AAV40819.1; -; Genomic_DNA. DR EMBL; LN614756; CEJ96915.1; -; Genomic_DNA. DR EMBL; LN831031; CKH72398.1; -; Genomic_DNA. DR RefSeq; WP_009888224.1; NZ_WUUI01000021.1. 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 AlphaFoldDB; Q5U924; -. DR SMR; Q5U924; -. DR GeneID; 66352922; -. DR KEGG; pdf:CD630DERM_03970; -. DR BRENDA; 4.2.1.157; 1473. DR SABIO-RK; Q5U924; -. DR EvolutionaryTrace; Q5U924; -. 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 Gene3D; 3.40.50.11890; -; 1. DR Gene3D; 3.40.50.11900; -; 1. DR InterPro; IPR010327; FldB/FldC_alpha/beta. DR PANTHER; PTHR30548; 2-HYDROXYGLUTARYL-COA DEHYDRATASE, D-COMPONENT-RELATED; 1. DR PANTHER; PTHR30548:SF4; SUBUNIT OF OXYGEN-SENSITIVE 2-HYDROXYISOCAPROYL-COA DEHYDRATASE; 1. DR Pfam; PF06050; HGD-D; 1. PE 1: Evidence at protein level; KW 3D-structure; 4Fe-4S; Direct protein sequencing; Iron; Iron-sulfur; Lyase; KW Metal-binding; Plasmid. FT INIT_MET 1 FT /note="Removed" FT /evidence="ECO:0000269|PubMed:15654892" FT CHAIN 2..408 FT /note="(R)-2-hydroxyisocaproyl-CoA dehydratase alpha FT subunit" FT /id="PRO_0000435662" FT BINDING 55 FT /ligand="substrate" FT /evidence="ECO:0000269|PubMed:21366233" FT BINDING 84 FT /ligand="[4Fe-4S] cluster" FT /ligand_id="ChEBI:CHEBI:49883" FT /evidence="ECO:0000269|PubMed:21366233" FT BINDING 117 FT /ligand="[4Fe-4S] cluster" FT /ligand_id="ChEBI:CHEBI:49883" FT /evidence="ECO:0000269|PubMed:21366233" FT BINDING 346 FT /ligand="[4Fe-4S] cluster" FT /ligand_id="ChEBI:CHEBI:49883" FT /evidence="ECO:0000269|PubMed:21366233" FT HELIX 7..28 FT /evidence="ECO:0007829|PDB:3O3M" FT STRAND 32..35 FT /evidence="ECO:0007829|PDB:3O3M" FT HELIX 42..45 FT /evidence="ECO:0007829|PDB:3O3M" FT TURN 46..48 FT /evidence="ECO:0007829|PDB:3O3M" FT STRAND 50..52 FT /evidence="ECO:0007829|PDB:3O3M" FT HELIX 54..63 FT /evidence="ECO:0007829|PDB:3O3M" FT HELIX 67..76 FT /evidence="ECO:0007829|PDB:3O3M" FT HELIX 85..96 FT /evidence="ECO:0007829|PDB:3O3M" FT STRAND 100..102 FT /evidence="ECO:0007829|PDB:3O3M" FT STRAND 108..113 FT /evidence="ECO:0007829|PDB:3O3M" FT HELIX 119..131 FT /evidence="ECO:0007829|PDB:3O3M" FT STRAND 135..138 FT /evidence="ECO:0007829|PDB:3O3M" FT STRAND 145..147 FT /evidence="ECO:0007829|PDB:3O3M" FT HELIX 150..171 FT /evidence="ECO:0007829|PDB:3O3M" FT HELIX 177..197 FT /evidence="ECO:0007829|PDB:3O3M" FT TURN 198..203 FT /evidence="ECO:0007829|PDB:3O3M" FT HELIX 211..213 FT /evidence="ECO:0007829|PDB:3O3M" FT HELIX 214..223 FT /evidence="ECO:0007829|PDB:3O3M" FT HELIX 228..246 FT /evidence="ECO:0007829|PDB:3O3M" FT STRAND 258..264 FT /evidence="ECO:0007829|PDB:3O3M" FT HELIX 268..270 FT /evidence="ECO:0007829|PDB:3O3M" FT HELIX 271..280 FT /evidence="ECO:0007829|PDB:3O3M" FT STRAND 283..287 FT /evidence="ECO:0007829|PDB:3O3M" FT HELIX 290..292 FT /evidence="ECO:0007829|PDB:3O3M" FT TURN 293..295 FT /evidence="ECO:0007829|PDB:3O3M" FT HELIX 303..311 FT /evidence="ECO:0007829|PDB:3O3M" FT HELIX 314..316 FT /evidence="ECO:0007829|PDB:3O3M" FT HELIX 319..332 FT /evidence="ECO:0007829|PDB:3O3M" FT STRAND 336..345 FT /evidence="ECO:0007829|PDB:3O3M" FT HELIX 347..350 FT /evidence="ECO:0007829|PDB:3O3M" FT HELIX 353..364 FT /evidence="ECO:0007829|PDB:3O3M" FT STRAND 368..373 FT /evidence="ECO:0007829|PDB:3O3M" FT HELIX 378..380 FT /evidence="ECO:0007829|PDB:3O3M" FT HELIX 383..401 FT /evidence="ECO:0007829|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 //