Reviewed, UniProtKB/Swiss-Prot P07445 (SDIS_PSEPU)

Names and origin

Protein names

Recommended name:
    Steroid Delta-isomerase
    EC=5.3.3.1
Alternative name(s):
    Delta(5)-3-ketosteroid isomerase

Gene names

Name:

ksi

Organism

Pseudomonas putida

Taxonomic identifier

303 [NCBI]

Taxonomic lineage

Bacteria › Proteobacteria › Gammaproteobacteria › Pseudomonadales › Pseudomonadaceae › Pseudomonas

Protein attributes

Sequence length	131 AA.
Sequence status	Complete.
Sequence processing	The displayed sequence is not processed.
Protein existence	Evidence at protein level.

General annotation (Comments)

Catalytic activity	A 3-oxo-Delta(5)-steroid = a 3-oxo-Delta(4)-steroid.
Subunit structure	Homodimer.
Induction	By steroids.

Ontologies

Keywords
Biological process	Lipid metabolism Steroid metabolism
Molecular function	Isomerase
Technical term	3D-structure Direct protein sequencing
Gene Ontology (GO)
Biological process	steroid metabolic process Inferred from electronic annotation. Source: UniProtKB-KW transport Inferred from electronic annotation. Source: InterPro
Cellular component	intracellular Inferred from electronic annotation. Source: InterPro
Molecular function	steroid delta-isomerase activity Inferred from electronic annotation. Source: EC
Complete GO annotation...

Sequence annotation (Features)

Feature key

Position(s)

Length

Description

Graphical view

Feature identifier

Molecule processing

Chain

1 – 131

131

Steroid Delta-isomerase

PRO_0000097646

Sites

Active site

16

1

Proton donor

Active site

40

1

Proton acceptor

Binding site

103

1

Substrate

Experimental info

Mutagenesis

16

1

Y → F: Reduces activity 2000-fold. Reduces activity 10000-fold; when associated with E-103; N-103 or L-103. Ref.3 Ref.5 Ref.7

Mutagenesis

16

1

Y → S: Reduces activity 20-fold. Ref.3 Ref.5 Ref.7

Mutagenesis

32

1

Y → S: Reduces activity 4-fold. Ref.7

Mutagenesis

57

1

Y → S: Reduces activity 100-fold.

Mutagenesis

92

1

W → A: Slightly reduces activity. Reduces protein stability.

Mutagenesis

103

1

D → A or L: Reduces activity 100-fold. Reduces activity 10000-fold; when associated with F-16. Ref.3 Ref.5

Mutagenesis

103

1

D → E: Slightly reduces activity. Reduces activity 10000-fold; when associated with F-16. Ref.3 Ref.5

Mutagenesis

103

1

D → N: Reduces activity 4-fold. Reduces activity 10000-fold; when associated with F-16. Ref.3 Ref.5

Mutagenesis

125

1

L → A: Slightly reduces activity and reduces protein stability; when associated with A-127. Ref.8

Mutagenesis

127

1

V → A: Slightly reduces activity and reduces protein stability; when associated with A-125. Ref.8

Secondary structure

1

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

.

131

Helix Strand Turn

Details...

Helix

6 – 22

17

Helix

25 – 31

7

Beta strand

32 – 41

10

Helix

49 – 61

13

Beta strand

62 – 64

3

Beta strand

67 – 72

6

Beta strand

78 – 107

30

Beta strand

113 – 119

7

Helix

122 – 124

3

Sequences

	Sequence		Length	Mass (Da)
	P07445-1 [UniParc]. Last modified April 1, 1988. Version 1. Checksum: 08711198C13CF014	FASTA	131	14,536
	10 20 30 40 50 60 MNLPTAQEVQ GLMARYIELV DVGDIEAIVQ MYADDATVED PFGQPPIHGR EQIAAFYRQG 70 80 90 100 110 120 LGGGKVRACL TGPVRASHNG CGAMPFRVEM VWNGQPCALD VIDVMRFDEH GRIQTMQAYW 130 SEVNLSVREP Q

References

[1]	"Cloning, nucleotide sequence, and overexpression of the gene coding for delta 5-3-ketosteroid isomerase from Pseudomonas putida biotype B." Kim S.W., Kim C.Y., Benisek W.F., Choi K.Y. J. Bacteriol. 176:6672-6676(1994) [PubMed: 7961420] [Abstract] Cited for: NUCLEOTIDE SEQUENCE [GENOMIC DNA]. Strain: Biotype B.
[2]	"The amino acid sequence of a delta 5-3-oxosteroid isomerase from Pseudomonas putida biotype B." Linden K.G., Benisek W.F. J. Biol. Chem. 261:6454-6460(1986) [PubMed: 3700400] [Abstract] Cited for: PROTEIN SEQUENCE. Strain: Biotype B.
[3]	"High-resolution crystal structures of delta5-3-ketosteroid isomerase with and without a reaction intermediate analogue." Kim S.-W., Cha S.-S., Cho H.-S., Kim J.-S., Ha N.-C., Cho M.-J., Joo S., Kim K.-K., Choi K.-Y., Oh B.-H. Biochemistry 36:14030-14036(1997) [PubMed: 9369474] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (2.5 ANGSTROMS) IN COMPLEX WITH REACTION INTERMEDIATE ANALOG, MUTAGENESIS OF TYR-16 AND ASP-103.
[4]	"Roles of active site aromatic residues in catalysis by ketosteroid isomerase from Pseudomonas putida biotype B." Kim D.-H., Nam G.H., Jang D.S., Choi G., Joo S., Kim J.-S., Oh B.-H., Choi K.-Y. Biochemistry 38:13810-13819(1999) [PubMed: 10529226] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (2.1 ANGSTROMS).
[5]	"Asp-99 donates a hydrogen bond not to Tyr-14 but to the steroid directly in the catalytic mechanism of delta 5-3-ketosteroid isomerase from Pseudomonas putida biotype B." Choi G., Ha N.-C., Kim S.W., Kim D.-H., Park S., Oh B.-H., Choi K.-Y. Biochemistry 39:903-909(2000) [PubMed: 10653633] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (1.9 ANGSTROMS) OF MUTANT ASN-40/GLU-103 IN COMPLEX WITH REACTION INTERMEDIATE ANALOG, MUTAGENESIS OF TYR-16 AND ASP-103.
[6]	"Detection of large pKa perturbations of an inhibitor and a catalytic group at an enzyme active site, a mechanistic basis for catalytic power of many enzymes." Ha N.-C., Kim M.-S., Lee W., Choi K.-Y., Oh B.-H. J. Biol. Chem. 275:41100-41106(2000) [PubMed: 11007792] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (2.5 ANGSTROMS) IN COMPLEX WITH SUBSTRATE.
[7]	"Maintenance of alpha-helical structures by phenyl rings in the active-site tyrosine triad contributes to catalysis and stability of ketosteroid isomerase from Pseudomonas putida biotype B." Nam G.H., Jang D.S., Cha S.-S., Lee T.-H., Kim D.-H., Hong B.H., Yun Y.S., Oh B.-H., Choi K.-Y. Biochemistry 40:13529-13537(2001) [PubMed: 11695900] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (2.2 ANGSTROMS) OF MUTANT SER-57, MUTAGENESIS OF TYR-16 AND TYR-32.
[8]	"Small exterior hydrophobic cluster contributes to conformational stability and steroid binding in ketosteroid isomerase from Pseudomonas putida biotype B." Yun Y.S., Nam G.H., Kim Y.-G., Oh B.-H., Choi K.-Y. FEBS J. 272:1999-2011(2005) [PubMed: 15819891] [Abstract] Cited for: X-RAY CRYSTALLOGRAPHY (2.1 ANGSTROMS) OF MUTANT ALA-92, MUTAGENESIS OF LEU-125 AND VAL-127.
+	Additional computationally mapped references.

Cross-references

Sequence databases

EMBLGenBankDDBJ

L13127 Genomic DNA. Translation: AAA64437.1.

PIR

SIPSDP. A25216.

3D structure databases

PDBPDBe

Entry	Method	Resolution (Å)	Chain	Positions	PDBsum
1C7H	X-ray	2.50	A	1-131	[»]
1CQS	X-ray	1.90	A/B	1-131	[»]
1DMM	X-ray	1.90	A	1-131	[»]
1DMN	X-ray	2.05	A	1-131	[»]
1DMQ	X-ray	2.15	A	1-131	[»]
1E3R	X-ray	2.50	A/B	1-131	[»]
1E3V	X-ray	2.00	A/B	1-131	[»]
1E97	X-ray	2.00	A	1-131	[»]
1EA2	X-ray	1.80	A	1-131	[»]
1GS3	X-ray	2.10	A	1-131	[»]
1K41	X-ray	2.20	A/B	1-131	[»]
1OGX	X-ray	2.00	A/B	1-131	[»]
1OH0	X-ray	1.10	A/B	1-131	[»]
1OHO	X-ray	1.90	A	1-131	[»]
1OPY	X-ray	1.90	A	1-131	[»]
1VZZ	X-ray	2.30	A/B	1-131	[»]
1W00	X-ray	2.20	A/B	1-131	[»]
1W01	X-ray	2.20	A/B	1-131	[»]
1W02	X-ray	2.30	A	1-131	[»]
1W6Y	X-ray	2.10	A	1-131	[»]
2INX	X-ray	1.50	A	1-131	[»]
2PZV	X-ray	1.25	A/B/C/D	1-131	[»]
3CPO	X-ray	1.24	A	1-131	[»]

ModBase

Search...

Enzyme and pathway databases

BRENDA

5.3.3.1. 403.

Family and domain databases

InterPro

IPR002075. NTF2.
[Graphical view]

Pfam

PF02136. NTF2. 1 hit.
[Graphical view]

ProtoNet

Search...

Entry information

Entry name

SDIS_PSEPU

Accession

Primary (citable) accession number: P07445

Entry history

Integrated into UniProtKB/Swiss-Prot:	April 1, 1988
Last sequence update:	April 1, 1988
Last modified:	June 16, 2009
This is version 72 of the entry and version 1 of the sequence. [Complete history]

Entry status

Reviewed (UniProtKB/Swiss-Prot)

Annotation project

HAMAP (High-quality Automated and Manual Annotation of microbial Proteomes)