ID SDIS_PSEPU Reviewed; 131 AA. AC P07445; DT 01-APR-1988, integrated into UniProtKB/Swiss-Prot. DT 01-APR-1988, sequence version 1. DT 27-MAR-2024, entry version 124. DE RecName: Full=Steroid Delta-isomerase; DE EC=5.3.3.1; DE AltName: Full=Delta(5)-3-ketosteroid isomerase; GN Name=ksi; OS Pseudomonas putida (Arthrobacter siderocapsulatus). OC Bacteria; Pseudomonadota; Gammaproteobacteria; Pseudomonadales; OC Pseudomonadaceae; Pseudomonas. OX NCBI_TaxID=303; RN [1] RP NUCLEOTIDE SEQUENCE [GENOMIC DNA]. RC STRAIN=Biotype B; RX PubMed=7961420; DOI=10.1128/jb.176.21.6672-6676.1994; RA Kim S.W., Kim C.Y., Benisek W.F., Choi K.Y.; RT "Cloning, nucleotide sequence, and overexpression of the gene coding for RT delta 5-3-ketosteroid isomerase from Pseudomonas putida biotype B."; RL J. Bacteriol. 176:6672-6676(1994). RN [2] RP PROTEIN SEQUENCE. RC STRAIN=Biotype B; RX PubMed=3700400; DOI=10.1016/s0021-9258(19)84584-2; RA Linden K.G., Benisek W.F.; RT "The amino acid sequence of a delta 5-3-oxosteroid isomerase from RT Pseudomonas putida biotype B."; RL J. Biol. Chem. 261:6454-6460(1986). RN [3] RP X-RAY CRYSTALLOGRAPHY (2.5 ANGSTROMS) IN COMPLEX WITH REACTION INTERMEDIATE RP ANALOG, AND MUTAGENESIS OF TYR-16 AND ASP-103. RX PubMed=9369474; DOI=10.1021/bi971546+; RA Kim S.-W., Cha S.-S., Cho H.-S., Kim J.-S., Ha N.-C., Cho M.-J., Joo S., RA Kim K.-K., Choi K.-Y., Oh B.-H.; RT "High-resolution crystal structures of delta5-3-ketosteroid isomerase with RT and without a reaction intermediate analogue."; RL Biochemistry 36:14030-14036(1997). RN [4] RP X-RAY CRYSTALLOGRAPHY (2.1 ANGSTROMS). RX PubMed=10529226; DOI=10.1021/bi991040m; RA Kim D.-H., Nam G.H., Jang D.S., Choi G., Joo S., Kim J.-S., Oh B.-H., RA Choi K.-Y.; RT "Roles of active site aromatic residues in catalysis by ketosteroid RT isomerase from Pseudomonas putida biotype B."; RL Biochemistry 38:13810-13819(1999). RN [5] RP X-RAY CRYSTALLOGRAPHY (1.9 ANGSTROMS) OF MUTANT ASN-40/GLU-103 IN COMPLEX RP WITH REACTION INTERMEDIATE ANALOG, AND MUTAGENESIS OF TYR-16 AND ASP-103. RX PubMed=10653633; DOI=10.1021/bi991579k; RA Choi G., Ha N.-C., Kim S.W., Kim D.-H., Park S., Oh B.-H., Choi K.-Y.; RT "Asp-99 donates a hydrogen bond not to Tyr-14 but to the steroid directly RT in the catalytic mechanism of delta 5-3-ketosteroid isomerase from RT Pseudomonas putida biotype B."; RL Biochemistry 39:903-909(2000). RN [6] RP X-RAY CRYSTALLOGRAPHY (2.5 ANGSTROMS) IN COMPLEX WITH SUBSTRATE. RX PubMed=11007792; DOI=10.1074/jbc.m007561200; RA Ha N.-C., Kim M.-S., Lee W., Choi K.-Y., Oh B.-H.; RT "Detection of large pKa perturbations of an inhibitor and a catalytic group RT at an enzyme active site, a mechanistic basis for catalytic power of many RT enzymes."; RL J. Biol. Chem. 275:41100-41106(2000). RN [7] RP X-RAY CRYSTALLOGRAPHY (2.2 ANGSTROMS) OF MUTANT SER-57, AND MUTAGENESIS OF RP TYR-16 AND TYR-32. RX PubMed=11695900; DOI=10.1021/bi015547k; RA Nam G.H., Jang D.S., Cha S.-S., Lee T.-H., Kim D.-H., Hong B.H., Yun Y.S., RA Oh B.-H., Choi K.-Y.; RT "Maintenance of alpha-helical structures by phenyl rings in the active-site RT tyrosine triad contributes to catalysis and stability of ketosteroid RT isomerase from Pseudomonas putida biotype B."; RL Biochemistry 40:13529-13537(2001). RN [8] RP X-RAY CRYSTALLOGRAPHY (2.1 ANGSTROMS) OF MUTANT ALA-92, AND MUTAGENESIS OF RP LEU-125 AND VAL-127. RX PubMed=15819891; DOI=10.1111/j.1742-4658.2005.04627.x; RA Yun Y.S., Nam G.H., Kim Y.-G., Oh B.-H., Choi K.-Y.; RT "Small exterior hydrophobic cluster contributes to conformational stability RT and steroid binding in ketosteroid isomerase from Pseudomonas putida RT biotype B."; RL FEBS J. 272:1999-2011(2005). CC -!- CATALYTIC ACTIVITY: CC Reaction=a 3-oxo-Delta(5)-steroid = a 3-oxo-Delta(4)-steroid; CC Xref=Rhea:RHEA:14709, ChEBI:CHEBI:47907, ChEBI:CHEBI:47909; CC EC=5.3.3.1; CC -!- SUBUNIT: Homodimer. {ECO:0000269|PubMed:10653633, CC ECO:0000269|PubMed:11007792, ECO:0000269|PubMed:9369474}. CC -!- INDUCTION: By steroids. 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; L13127; AAA64437.1; -; Genomic_DNA. DR PIR; A25216; SIPSDP. DR PDB; 1C7H; X-ray; 2.50 A; A=1-131. DR PDB; 1CQS; X-ray; 1.90 A; A/B=1-131. DR PDB; 1DMM; X-ray; 1.90 A; A=1-131. DR PDB; 1DMN; X-ray; 2.05 A; A=1-131. DR PDB; 1DMQ; X-ray; 2.15 A; A=1-131. DR PDB; 1E3R; X-ray; 2.50 A; A/B=1-131. DR PDB; 1E3V; X-ray; 2.00 A; A/B=1-131. DR PDB; 1E97; X-ray; 2.00 A; A=1-131. DR PDB; 1EA2; X-ray; 1.80 A; A=1-131. DR PDB; 1GS3; X-ray; 2.10 A; A=1-131. DR PDB; 1K41; X-ray; 2.20 A; A/B=1-131. DR PDB; 1OGX; X-ray; 2.00 A; A/B=1-131. DR PDB; 1OH0; X-ray; 1.10 A; A/B=1-131. DR PDB; 1OHO; X-ray; 1.90 A; A=1-131. DR PDB; 1OPY; X-ray; 1.90 A; A=1-131. DR PDB; 1VZZ; X-ray; 2.30 A; A/B=1-131. DR PDB; 1W00; X-ray; 2.20 A; A/B=1-131. DR PDB; 1W01; X-ray; 2.20 A; A/B=1-131. DR PDB; 1W02; X-ray; 2.30 A; A=1-131. DR PDB; 1W6Y; X-ray; 2.10 A; A=1-131. DR PDB; 2INX; X-ray; 1.50 A; A=1-131. DR PDB; 2PZV; X-ray; 1.25 A; A/B/C/D=1-131. DR PDB; 3CPO; X-ray; 1.24 A; A=1-131. DR PDB; 3FZW; X-ray; 1.32 A; A/B=1-131. DR PDB; 3IPT; X-ray; 1.63 A; A/B/C/D=1-131. DR PDB; 3OWS; X-ray; 1.71 A; A/B/C/D=1-131. DR PDB; 3OWU; X-ray; 1.70 A; A/B/C/D=1-131. DR PDB; 3OWY; X-ray; 2.30 A; A/B/C/D/E/F/G/H=1-131. DR PDB; 3OX9; X-ray; 2.00 A; A/B/C/D=1-131. DR PDB; 3OXA; X-ray; 1.89 A; A/B/C/D=1-131. DR PDB; 3RGR; X-ray; 1.59 A; A=1-131. DR PDB; 3SED; X-ray; 1.30 A; A=3-127. DR PDB; 3T8N; X-ray; 1.47 A; A/B/D/F=1-131. DR PDB; 3VGN; X-ray; 1.30 A; A/B=1-131. DR PDB; 3VSY; X-ray; 1.50 A; A/B=3-131. DR PDB; 4CDL; X-ray; 2.50 A; A=1-131. DR PDB; 4K1U; X-ray; 2.00 A; A/B=1-131. DR PDB; 4K1V; X-ray; 1.80 A; A=1-131. DR PDB; 5AI1; X-ray; 2.10 A; A=1-131. DR PDB; 5D81; X-ray; 1.39 A; A=1-131. DR PDB; 5D82; X-ray; 1.37 A; A/B=1-131. DR PDB; 5D83; X-ray; 1.70 A; A/B=1-131. DR PDB; 5G2G; X-ray; 1.60 A; A/B=2-128. DR PDB; 5KP1; X-ray; 1.22 A; A/B/C/D=1-131. DR PDB; 5KP3; X-ray; 1.70 A; A/B=1-131. DR PDB; 5KP4; X-ray; 1.71 A; A/B=1-131. DR PDB; 6C17; X-ray; 1.10 A; A=1-131. DR PDB; 6C1J; X-ray; 1.06 A; A=1-131. DR PDB; 6C1X; X-ray; 1.05 A; A=1-131. DR PDB; 6F4Y; X-ray; 1.92 A; A/B=3-127. DR PDB; 6F50; X-ray; 2.00 A; A/B=3-127. DR PDB; 6F53; X-ray; 1.49 A; A=3-127. DR PDB; 6F54; X-ray; 1.08 A; A/B=3-127. DR PDB; 6TZD; X-ray; 1.45 A; A/B=1-131. DR PDB; 6U1Z; X-ray; 1.50 A; A/B=1-131. DR PDB; 6U4I; X-ray; 1.55 A; A/B=1-131. DR PDB; 6UBQ; X-ray; 1.30 A; A/B=1-131. DR PDB; 6UCN; X-ray; 1.32 A; A/B=1-131. DR PDB; 6UCW; X-ray; 1.25 A; A/B=1-131. DR PDB; 6UCY; X-ray; 1.15 A; A/B=1-131. DR PDB; 6UFS; X-ray; 1.47 A; A/B=1-131. DR PDB; 7RXF; X-ray; 1.16 A; A/B=1-131. DR PDB; 7RXK; X-ray; 1.10 A; A/B=1-131. DR PDB; 7RY4; X-ray; 1.11 A; A/B=1-131. DR PDBsum; 1C7H; -. DR PDBsum; 1CQS; -. DR PDBsum; 1DMM; -. DR PDBsum; 1DMN; -. DR PDBsum; 1DMQ; -. DR PDBsum; 1E3R; -. DR PDBsum; 1E3V; -. DR PDBsum; 1E97; -. DR PDBsum; 1EA2; -. DR PDBsum; 1GS3; -. DR PDBsum; 1K41; -. DR PDBsum; 1OGX; -. DR PDBsum; 1OH0; -. DR PDBsum; 1OHO; -. DR PDBsum; 1OPY; -. DR PDBsum; 1VZZ; -. DR PDBsum; 1W00; -. DR PDBsum; 1W01; -. DR PDBsum; 1W02; -. DR PDBsum; 1W6Y; -. DR PDBsum; 2INX; -. DR PDBsum; 2PZV; -. DR PDBsum; 3CPO; -. DR PDBsum; 3FZW; -. DR PDBsum; 3IPT; -. DR PDBsum; 3OWS; -. DR PDBsum; 3OWU; -. DR PDBsum; 3OWY; -. DR PDBsum; 3OX9; -. DR PDBsum; 3OXA; -. DR PDBsum; 3RGR; -. DR PDBsum; 3SED; -. DR PDBsum; 3T8N; -. DR PDBsum; 3VGN; -. DR PDBsum; 3VSY; -. DR PDBsum; 4CDL; -. DR PDBsum; 4K1U; -. DR PDBsum; 4K1V; -. DR PDBsum; 5AI1; -. DR PDBsum; 5D81; -. DR PDBsum; 5D82; -. DR PDBsum; 5D83; -. DR PDBsum; 5G2G; -. DR PDBsum; 5KP1; -. DR PDBsum; 5KP3; -. DR PDBsum; 5KP4; -. DR PDBsum; 6C17; -. DR PDBsum; 6C1J; -. DR PDBsum; 6C1X; -. DR PDBsum; 6F4Y; -. DR PDBsum; 6F50; -. DR PDBsum; 6F53; -. DR PDBsum; 6F54; -. DR PDBsum; 6TZD; -. DR PDBsum; 6U1Z; -. DR PDBsum; 6U4I; -. DR PDBsum; 6UBQ; -. DR PDBsum; 6UCN; -. DR PDBsum; 6UCW; -. DR PDBsum; 6UCY; -. DR PDBsum; 6UFS; -. DR PDBsum; 7RXF; -. DR PDBsum; 7RXK; -. DR PDBsum; 7RY4; -. DR AlphaFoldDB; P07445; -. DR SMR; P07445; -. DR BindingDB; P07445; -. DR ChEMBL; CHEMBL2321641; -. DR DrugBank; DB03619; Deoxycholic acid. DR DrugBank; DB03515; Equilenin. DR DrugCentral; P07445; -. DR BRENDA; 5.3.3.1; 5092. DR EvolutionaryTrace; P07445; -. DR GO; GO:0004769; F:steroid delta-isomerase activity; IEA:UniProtKB-EC. DR GO; GO:0008202; P:steroid metabolic process; IEA:UniProtKB-KW. DR CDD; cd00781; ketosteroid_isomerase; 1. DR Gene3D; 3.10.450.50; -; 1. DR InterPro; IPR039256; Ketosteroid_isomerase. DR InterPro; IPR032710; NTF2-like_dom_sf. DR InterPro; IPR037401; SnoaL-like. DR Pfam; PF12680; SnoaL_2; 1. DR SUPFAM; SSF54427; NTF2-like; 1. PE 1: Evidence at protein level; KW 3D-structure; Direct protein sequencing; Isomerase; Lipid metabolism; KW Steroid metabolism. FT CHAIN 1..131 FT /note="Steroid Delta-isomerase" FT /id="PRO_0000097646" FT ACT_SITE 16 FT /note="Proton donor" FT ACT_SITE 40 FT /note="Proton acceptor" FT BINDING 103 FT /ligand="substrate" FT /evidence="ECO:0000269|PubMed:11007792" FT MUTAGEN 16 FT /note="Y->F: Reduces activity 2000-fold. Reduces activity FT 10000-fold; when associated with E-103; N-103 or L-103." FT /evidence="ECO:0000269|PubMed:10653633, FT ECO:0000269|PubMed:11695900, ECO:0000269|PubMed:9369474" FT MUTAGEN 16 FT /note="Y->S: Reduces activity 20-fold." FT /evidence="ECO:0000269|PubMed:10653633, FT ECO:0000269|PubMed:11695900, ECO:0000269|PubMed:9369474" FT MUTAGEN 32 FT /note="Y->S: Reduces activity 4-fold." FT /evidence="ECO:0000269|PubMed:11695900" FT MUTAGEN 57 FT /note="Y->S: Reduces activity 100-fold." FT MUTAGEN 92 FT /note="W->A: Slightly reduces activity. Reduces protein FT stability." FT MUTAGEN 103 FT /note="D->A,L: Reduces activity 100-fold. Reduces activity FT 10000-fold; when associated with F-16." FT /evidence="ECO:0000269|PubMed:10653633, FT ECO:0000269|PubMed:9369474" FT MUTAGEN 103 FT /note="D->E: Slightly reduces activity. Reduces activity FT 10000-fold; when associated with F-16." FT /evidence="ECO:0000269|PubMed:10653633, FT ECO:0000269|PubMed:9369474" FT MUTAGEN 103 FT /note="D->N: Reduces activity 4-fold. Reduces activity FT 10000-fold; when associated with F-16." FT /evidence="ECO:0000269|PubMed:10653633, FT ECO:0000269|PubMed:9369474" FT MUTAGEN 125 FT /note="L->A: Slightly reduces activity and reduces protein FT stability; when associated with A-127." FT /evidence="ECO:0000269|PubMed:15819891" FT MUTAGEN 127 FT /note="V->A: Slightly reduces activity and reduces protein FT stability; when associated with A-125." FT /evidence="ECO:0000269|PubMed:15819891" FT HELIX 6..22 FT /evidence="ECO:0007829|PDB:6C1X" FT HELIX 25..30 FT /evidence="ECO:0007829|PDB:6C1X" FT STRAND 32..41 FT /evidence="ECO:0007829|PDB:6C1X" FT HELIX 49..61 FT /evidence="ECO:0007829|PDB:6C1X" FT STRAND 62..64 FT /evidence="ECO:0007829|PDB:6C1J" FT STRAND 67..72 FT /evidence="ECO:0007829|PDB:6C1X" FT STRAND 78..92 FT /evidence="ECO:0007829|PDB:6C1X" FT STRAND 95..107 FT /evidence="ECO:0007829|PDB:6C1X" FT STRAND 111..119 FT /evidence="ECO:0007829|PDB:6C1X" FT HELIX 122..124 FT /evidence="ECO:0007829|PDB:6C1X" FT STRAND 125..127 FT /evidence="ECO:0007829|PDB:7RXK" SQ SEQUENCE 131 AA; 14536 MW; 08711198C13CF014 CRC64; MNLPTAQEVQ GLMARYIELV DVGDIEAIVQ MYADDATVED PFGQPPIHGR EQIAAFYRQG LGGGKVRACL TGPVRASHNG CGAMPFRVEM VWNGQPCALD VIDVMRFDEH GRIQTMQAYW SEVNLSVREP Q //