ID A0A2E9CYH6_9ACTN Unreviewed; 542 AA. AC A0A2E9CYH6; DT 31-JAN-2018, integrated into UniProtKB/TrEMBL. DT 31-JAN-2018, sequence version 1. DT 03-AUG-2022, entry version 21. DE RecName: Full=Multifunctional fusion protein {ECO:0000256|HAMAP-Rule:MF_00013, ECO:0000256|HAMAP-Rule:MF_00206}; DE Includes: DE RecName: Full=Lipoyl synthase {ECO:0000256|HAMAP-Rule:MF_00206}; DE EC=2.8.1.8 {ECO:0000256|HAMAP-Rule:MF_00206}; DE AltName: Full=Lip-syn {ECO:0000256|HAMAP-Rule:MF_00206}; DE AltName: Full=Lipoate synthase {ECO:0000256|HAMAP-Rule:MF_00206}; DE AltName: Full=Lipoic acid synthase {ECO:0000256|HAMAP-Rule:MF_00206}; DE AltName: Full=Sulfur insertion protein LipA {ECO:0000256|HAMAP-Rule:MF_00206}; DE Short=LS {ECO:0000256|HAMAP-Rule:MF_00206}; DE Includes: DE RecName: Full=Octanoyltransferase {ECO:0000256|HAMAP-Rule:MF_00013}; DE EC=2.3.1.181 {ECO:0000256|HAMAP-Rule:MF_00013}; DE AltName: Full=Lipoate-protein ligase B {ECO:0000256|HAMAP-Rule:MF_00013}; DE AltName: Full=Lipoyl/octanoyl transferase {ECO:0000256|HAMAP-Rule:MF_00013}; DE AltName: Full=Octanoyl-[acyl-carrier-protein]-protein N-octanoyltransferase {ECO:0000256|HAMAP-Rule:MF_00013}; GN Name=lipA {ECO:0000256|HAMAP-Rule:MF_00206}; GN Synonyms=lipB {ECO:0000256|HAMAP-Rule:MF_00013}; GN ORFNames=CL511_00020 {ECO:0000313|EMBL:MBS92057.1}; OS Actinomycetia bacterium. OC Bacteria; Actinobacteria. OX NCBI_TaxID=1883427 {ECO:0000313|EMBL:MBS92057.1, ECO:0000313|Proteomes:UP000230398}; RN [1] {ECO:0000313|Proteomes:UP000230398} RP NUCLEOTIDE SEQUENCE [LARGE SCALE GENOMIC DNA]. RA Tully B.J., Graham E.D., Heidelberg J.F.; RT "The Reconstruction of 2,631 Draft Metagenome-Assembled Genomes from the RT Global Oceans."; RL Submitted (SEP-2017) to the EMBL/GenBank/DDBJ databases. CC -!- FUNCTION: Catalyzes the radical-mediated insertion of two sulfur atoms CC into the C-6 and C-8 positions of the octanoyl moiety bound to the CC lipoyl domains of lipoate-dependent enzymes, thereby converting the CC octanoylated domains into lipoylated derivatives. {ECO:0000256|HAMAP- CC Rule:MF_00206}. CC -!- FUNCTION: Catalyzes the transfer of endogenously produced octanoic acid CC from octanoyl-acyl-carrier-protein onto the lipoyl domains of lipoate- CC dependent enzymes. Lipoyl-ACP can also act as a substrate although CC octanoyl-ACP is likely to be the physiological substrate. CC {ECO:0000256|ARBA:ARBA00024732, ECO:0000256|HAMAP-Rule:MF_00013}. CC -!- CATALYTIC ACTIVITY: CC Reaction=L-lysyl-[protein] + octanoyl-[ACP] = H(+) + holo-[ACP] + N(6)- CC octanoyl-L-lysyl-[protein]; Xref=Rhea:RHEA:17665, Rhea:RHEA- CC COMP:9636, Rhea:RHEA-COMP:9685, Rhea:RHEA-COMP:9752, Rhea:RHEA- CC COMP:9928, ChEBI:CHEBI:15378, ChEBI:CHEBI:29969, ChEBI:CHEBI:64479, CC ChEBI:CHEBI:78463, ChEBI:CHEBI:78809; EC=2.3.1.181; CC Evidence={ECO:0000256|ARBA:ARBA00000953, ECO:0000256|HAMAP- CC Rule:MF_00013}; CC -!- CATALYTIC ACTIVITY: CC Reaction=[[Fe-S] cluster scaffold protein carrying a second [4Fe- CC 4S](2+) cluster] + 4 H(+) + N(6)-octanoyl-L-lysyl-[protein] + 2 CC oxidized [2Fe-2S]-[ferredoxin] + 2 S-adenosyl-L-methionine = (R)- CC N(6)-dihydrolipoyl-L-lysyl-[protein] + 2 5'-deoxyadenosine + [[Fe-S] CC cluster scaffold protein] + 4 Fe(3+) + 2 hydrogen sulfide + 2 L- CC methionine + 2 reduced [2Fe-2S]-[ferredoxin]; Xref=Rhea:RHEA:16585, CC Rhea:RHEA-COMP:9928, Rhea:RHEA-COMP:10000, Rhea:RHEA-COMP:10001, CC Rhea:RHEA-COMP:10475, Rhea:RHEA-COMP:14568, Rhea:RHEA-COMP:14569, CC ChEBI:CHEBI:15378, ChEBI:CHEBI:17319, ChEBI:CHEBI:29034, CC ChEBI:CHEBI:29919, ChEBI:CHEBI:33722, ChEBI:CHEBI:33737, CC ChEBI:CHEBI:33738, ChEBI:CHEBI:57844, ChEBI:CHEBI:59789, CC ChEBI:CHEBI:78809, ChEBI:CHEBI:83100; EC=2.8.1.8; CC Evidence={ECO:0000256|ARBA:ARBA00001264, ECO:0000256|HAMAP- CC Rule:MF_00206}; CC -!- COFACTOR: CC Name=[4Fe-4S] cluster; Xref=ChEBI:CHEBI:49883; CC Evidence={ECO:0000256|HAMAP-Rule:MF_00206}; CC Note=Binds 2 [4Fe-4S] clusters per subunit. One cluster is coordinated CC with 3 cysteines and an exchangeable S-adenosyl-L-methionine. CC {ECO:0000256|HAMAP-Rule:MF_00206}; CC -!- PATHWAY: Protein modification; protein lipoylation via endogenous CC pathway; protein N(6)-(lipoyl)lysine from octanoyl-[acyl-carrier- CC protein]: step 1/2. {ECO:0000256|ARBA:ARBA00004821, ECO:0000256|HAMAP- CC Rule:MF_00013}. CC -!- PATHWAY: Protein modification; protein lipoylation via endogenous CC pathway; protein N(6)-(lipoyl)lysine from octanoyl-[acyl-carrier- CC protein]: step 2/2. {ECO:0000256|HAMAP-Rule:MF_00206}. CC -!- SUBCELLULAR LOCATION: Cytoplasm {ECO:0000256|HAMAP-Rule:MF_00013}. CC -!- MISCELLANEOUS: In the reaction, the free carboxyl group of octanoic CC acid is attached via an amide linkage to the epsilon-amino group of a CC specific lysine residue of lipoyl domains of lipoate-dependent enzymes. CC {ECO:0000256|HAMAP-Rule:MF_00013}. CC -!- SIMILARITY: Belongs to the LipB family. {ECO:0000256|HAMAP- CC Rule:MF_00013}. CC -!- SIMILARITY: Belongs to the radical SAM superfamily. Lipoyl synthase CC family. {ECO:0000256|HAMAP-Rule:MF_00206}. CC -!- CAUTION: Lacks conserved residue(s) required for the propagation of CC feature annotation. {ECO:0000256|HAMAP-Rule:MF_00206}. CC -!- CAUTION: The sequence shown here is derived from an EMBL/GenBank/DDBJ CC whole genome shotgun (WGS) entry which is preliminary data. CC {ECO:0000313|EMBL:MBS92057.1}. 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; PBXU01000001; MBS92057.1; -; Genomic_DNA. DR UniPathway; UPA00538; UER00592. DR Proteomes; UP000230398; Unassembled WGS sequence. DR GO; GO:0005737; C:cytoplasm; IEA:UniProtKB-SubCell. DR GO; GO:0051539; F:4 iron, 4 sulfur cluster binding; IEA:UniProtKB-UniRule. DR GO; GO:0016992; F:lipoate synthase activity; IEA:UniProtKB-UniRule. DR GO; GO:0102552; F:lipoyl synthase activity (acting on glycine-cleavage complex H protein; IEA:UniProtKB-EC. DR GO; GO:0102553; F:lipoyl synthase activity (acting on pyruvate dehydrogenase E2 protein); IEA:UniProtKB-EC. DR GO; GO:0033819; F:lipoyl(octanoyl) transferase activity; IEA:UniProtKB-EC. DR GO; GO:0046872; F:metal ion binding; IEA:UniProtKB-KW. DR GO; GO:0009249; P:protein lipoylation; IEA:UniProtKB-UniRule. DR CDD; cd16444; LipB; 1. DR Gene3D; 3.20.20.70; -; 1. DR Gene3D; 3.30.930.10; -; 1. DR HAMAP; MF_00013; LipB; 1. DR HAMAP; MF_00206; Lipoyl_synth; 1. DR InterPro; IPR045864; aa-tRNA-synth_II/BPL/LPL. DR InterPro; IPR013785; Aldolase_TIM. DR InterPro; IPR004143; BPL_LPL_catalytic. DR InterPro; IPR006638; Elp3/MiaB/NifB. DR InterPro; IPR031691; LIAS_N. DR InterPro; IPR003698; Lipoyl_synth. DR InterPro; IPR000544; Octanoyltransferase. DR InterPro; IPR020605; Octanoyltransferase_CS. DR InterPro; IPR007197; rSAM. DR PANTHER; PTHR10949; PTHR10949; 1. DR Pfam; PF16881; LIAS_N; 1. DR Pfam; PF04055; Radical_SAM; 1. DR SMART; SM00729; Elp3; 1. DR SUPFAM; SSF55681; SSF55681; 1. DR TIGRFAMs; TIGR00510; lipA; 1. DR TIGRFAMs; TIGR00214; lipB; 1. DR PROSITE; PS51733; BPL_LPL_CATALYTIC; 1. DR PROSITE; PS01313; LIPB; 1. DR PROSITE; PS51918; RADICAL_SAM; 1. PE 3: Inferred from homology; KW 4Fe-4S {ECO:0000256|HAMAP-Rule:MF_00206}; KW Acyltransferase {ECO:0000256|ARBA:ARBA00023315, ECO:0000256|HAMAP- KW Rule:MF_00013}; Cytoplasm {ECO:0000256|HAMAP-Rule:MF_00013}; KW Iron {ECO:0000256|ARBA:ARBA00023004, ECO:0000256|HAMAP-Rule:MF_00206}; KW Iron-sulfur {ECO:0000256|ARBA:ARBA00023014, ECO:0000256|HAMAP- KW Rule:MF_00206}; KW Metal-binding {ECO:0000256|ARBA:ARBA00022723, ECO:0000256|HAMAP- KW Rule:MF_00206}; KW S-adenosyl-L-methionine {ECO:0000256|ARBA:ARBA00022691, ECO:0000256|HAMAP- KW Rule:MF_00206}; KW Transferase {ECO:0000256|ARBA:ARBA00022679, ECO:0000256|HAMAP- KW Rule:MF_00013}. FT DOMAIN 32..212 FT /note="BPL/LPL catalytic" FT /evidence="ECO:0000259|PROSITE:PS51733" FT DOMAIN 305..521 FT /note="Radical SAM core" FT /evidence="ECO:0000259|PROSITE:PS51918" FT REGION 77..84 FT /note="Substrate binding" FT /evidence="ECO:0000256|HAMAP-Rule:MF_00013" FT REGION 143..145 FT /note="Substrate binding" FT /evidence="ECO:0000256|HAMAP-Rule:MF_00013" FT REGION 156..158 FT /note="Substrate binding" FT /evidence="ECO:0000256|HAMAP-Rule:MF_00013" FT ACT_SITE 174 FT /note="Acyl-thioester intermediate" FT /evidence="ECO:0000256|HAMAP-Rule:MF_00013" FT BINDING 293 FT /ligand="[4Fe-4S] cluster" FT /ligand_id="ChEBI:CHEBI:49883" FT /ligand_label="1" FT /evidence="ECO:0000256|HAMAP-Rule:MF_00206" FT BINDING 298 FT /ligand="[4Fe-4S] cluster" FT /ligand_id="ChEBI:CHEBI:49883" FT /ligand_label="1" FT /evidence="ECO:0000256|HAMAP-Rule:MF_00206" FT BINDING 304 FT /ligand="[4Fe-4S] cluster" FT /ligand_id="ChEBI:CHEBI:49883" FT /ligand_label="1" FT /evidence="ECO:0000256|HAMAP-Rule:MF_00206" FT BINDING 323 FT /ligand="[4Fe-4S] cluster" FT /ligand_id="ChEBI:CHEBI:49883" FT /ligand_label="2" FT /ligand_note="4Fe-4S-S-AdoMet" FT /evidence="ECO:0000256|HAMAP-Rule:MF_00206" FT BINDING 326 FT /ligand="[4Fe-4S] cluster" FT /ligand_id="ChEBI:CHEBI:49883" FT /ligand_label="2" FT /ligand_note="4Fe-4S-S-AdoMet" FT /evidence="ECO:0000256|HAMAP-Rule:MF_00206" FT BINDING 532 FT /ligand="[4Fe-4S] cluster" FT /ligand_id="ChEBI:CHEBI:49883" FT /ligand_label="1" FT /evidence="ECO:0000256|HAMAP-Rule:MF_00206" FT SITE 140 FT /note="Lowers pKa of active site Cys" FT /evidence="ECO:0000256|HAMAP-Rule:MF_00013" SQ SEQUENCE 542 AA; 60946 MW; 0D2E5CA28B5F1C8D CRC64; MRDLNIRWLG KLPYSEAYDL QLGLHRSVSQ EDSKDDYLLL LEHNNVITSG RSSKENNLLV SKGQLHELGI EYFETDRGGD ITYHGDGQLI GYPIIRLSDP KKVIPFVRNL ENVIIDSLRK FKIDSFTKED DTGVWTAKGK IASVGIKVSK WTTYHGFSLN IFDSLDGYQL INPCGNQSEQ ITSIHQFNPD ISFEEVASEI SDNFVKVFGY ANTDRQFSQF TPRQLKRTKE FNIDQMVKDG VFKINQNKIP VTVRGVLPSE PKRPEWMKVK ANLGSDYVSL KNLLSEKKLN TVCEEASCPN IYECWSMGTA TFMIMGDVXT RACGFCDVKT GRPGELDLGE PLRVAESVXA MNLTHAVITS VNRDDLEDGG SMFFADTIRA VKDKNSHCDV EVLVPDFKGL RSAIQNIIDA SPEVFNHNLE TVPRLQREIR TAASYGRSLS LLEYVKKQGF MGKTKTGLIV GMGETKEEVI SVLKDLSKIE VDIVTIGQYL RPTAKHRPID RYATIEEFED YKIIGESYGI PHVESGPLVR SSYHAKDSFA SA //