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Q7SIG4 (DFPA_LOLVU) Reviewed, UniProtKB/Swiss-Prot

Last modified July 9, 2014. Version 50. Feed History...

Clusters with 100%, 90%, 50% identity | Documents (1) | Third-party data text xml rdf/xml gff fasta
to top of pageNames·Attributes·General annotation·Ontologies·Sequence annotation·Sequences·References·Cross-refs·Entry info·DocumentsCustomize order

Names and origin

Protein namesRecommended name:
Diisopropyl-fluorophosphatase

Short name=DFPase
EC=3.1.8.2
OrganismLoligo vulgaris (Common European squid)
Taxonomic identifier6622 [NCBI]
Taxonomic lineageEukaryotaMetazoaLophotrochozoaMolluscaCephalopodaColeoideaNeocoleoideaDecapodiformesTeuthidaMyopsinaLoliginidaeLoligo

Protein attributes

Sequence length314 AA.
Sequence statusComplete.
Protein existenceEvidence at protein level

General annotation (Comments)

Function

Biological function and substrate unknown. However, it is capable of acting on phosphorus anhydride bonds (such as phosphorus-halide and phosphorus-cyanide) in organophosphorus compounds (including nerve gases). Ref.3

Catalytic activity

Diisopropyl fluorophosphate + H2O = diisopropyl phosphate + fluoride. Ref.1 Ref.2

Cofactor

Binds 2 calcium ions per subunit. Ref.2 Ref.5 Ref.6

Enzyme regulation

Inhibited by chelating agents. Ref.2

Subunit structure

Monomer. Ref.5 Ref.6

Biotechnological use

Has potential application in bio-remediation by detoxification of organo-phosphates used in insecticides or nerve agents used in chemical warfare such as soman, tabun and sarin. Ref.4

Biophysicochemical properties

Kinetic parameters:

KM=0.54 mM for diisopropyl-fluorophosphate (in the presence of 10 mM NaCl) Ref.1

KM=0.42 mM for diisopropyl-fluorophosphate (in the presence of 100 mM NaCl)

KM=0.34 mM for diisopropyl-fluorophosphate (in the presence of 200 mM NaCl)

KM=0.25 mM for diisopropyl-fluorophosphate (in the presence of 500 mM NaCl)

KM=0.17 mM for diisopropyl-fluorophosphate (in the presence of 1000 mM NaCl)

Vmax=130 µmol/min/mg enzyme (in the presence of 10 mM NaCl)

Vmax=170 µmol/min/mg enzyme (in the presence of 100 mM NaCl)

Vmax=307 µmol/min/mg enzyme (in the presence of 200 mM NaCl)

Vmax=326 µmol/min/mg enzyme (in the presence of 500 mM NaCl)

Vmax=214 µmol/min/mg enzyme (in the presence of 1000 mM NaCl)

pH dependence:

Optimum pH is 8.0. Ref.1

Temperature dependence:

Optimum temperature is 35 degrees Celsius. Ref.1

Ontologies

Keywords
   LigandCalcium
Metal-binding
   Molecular functionHydrolase
   Technical term3D-structure
Gene Ontology (GO)
   Biological_processmetabolic process

Inferred from direct assay Ref.3. Source: GOC

   Molecular_functioncalcium ion binding

Inferred from direct assay Ref.4. Source: UniProtKB

diisopropyl-fluorophosphatase activity

Inferred from direct assay Ref.3. Source: UniProtKB

Complete GO annotation...

Sequence annotation (Features)

Feature keyPosition(s)LengthDescriptionGraphical viewFeature identifier

Molecule processing

Chain1 – 314314Diisopropyl-fluorophosphatase
PRO_0000248834

Sites

Active site2871Proton acceptor Probable Ref.3
Metal binding211Calcium 1; catalytic Probable Ref.5 Ref.6
Metal binding1201Calcium 1; catalytic Probable Ref.5 Ref.6
Metal binding1751Calcium 1; catalytic Probable Ref.5 Ref.6
Metal binding2291Calcium 1; catalytic Probable Ref.5 Ref.6
Metal binding2321Calcium 2 Ref.5 Ref.6
Metal binding2731Calcium 2; via carbonyl oxygen Ref.5 Ref.6
Metal binding2741Calcium 2 Ref.5 Ref.6

Experimental info

Mutagenesis211E → Q: 100% decrease in activity. Loss of calcium 1 binding. Ref.5
Mutagenesis371E → Q: 50% decrease in activity. Ref.5
Mutagenesis771Q → F: 100% decrease in activity. Ref.3
Mutagenesis771Q → W: No effect on activity. Ref.3
Mutagenesis771Q → Y: 6% increase in activity. Ref.3
Mutagenesis1201N → D: 96% decrease in activity. 100% decrease in activity; when associated with N-229. Ref.3
Mutagenesis1211D → F: 100% decrease in activity. Ref.3
Mutagenesis1441Y → S: 8% increase in activity. Ref.3
Mutagenesis1461R → S: 45% decrease in activity. Ref.3
Mutagenesis1481M → A: 26% decrease in activity. Ref.3
Mutagenesis1731F → A: 84% decrease in activity. Ref.3
Mutagenesis1731F → L: 28% decrease in activity. Ref.3
Mutagenesis1731F → S: 68% decrease in activity. Ref.3
Mutagenesis1731F → V: 46% decrease in activity. Ref.3
Mutagenesis1731F → W: 19% decrease in activity. Ref.3
Mutagenesis1731F → Y: 53% decrease in activity. Ref.3
Mutagenesis1751N → D: 98% decrease in activity. Ref.3
Mutagenesis1811H → N: 20% decrease in activity. Ref.1 Ref.3
Mutagenesis1951T → A: 60% decrease in activity. Ref.3
Mutagenesis1951T → L: 11% decrease in activity. Ref.3
Mutagenesis1951T → V: 3% decrease in activity. Ref.3
Mutagenesis2191H → N: 3% increase in activity. Ref.1
Mutagenesis2241H → N: 14% increase in activity. Ref.1
Mutagenesis2291D → N: 100% decrease in activity. Loss of calcium 1 binding. 100% decrease in activity; when associated with D-120. Ref.3 Ref.5
Mutagenesis2321D → S: 3% increase in activity. 19% decrease in activity; when associated with A-271. Ref.3
Mutagenesis2371N → S: 4% decrease in activity. Ref.3
Mutagenesis2441W → F: 44% decrease in activity. Ref.5
Mutagenesis2441W → H: 27% decrease in activity. Ref.5
Mutagenesis2441W → L: 62% decrease in activity. Ref.5
Mutagenesis2441W → Y: No effect on activity. Ref.5
Mutagenesis2481H → N: 4% increase in activity. Ref.1
Mutagenesis2711S → A: 30% increase in activity. 19% decrease in activity; when associated with S-232. Ref.3 Ref.5
Mutagenesis2721N → F: 100% decrease in activity. Ref.3
Mutagenesis2741H → N: 85% decrease in activity. Ref.1 Ref.3
Mutagenesis2871H → A: 90% decrease in activity. Ref.1 Ref.3
Mutagenesis2871H → F: 36% decrease in activity. Ref.1 Ref.3
Mutagenesis2871H → L: 21% decrease in activity. Ref.1 Ref.3
Mutagenesis2871H → N: 97% decrease in activity. Ref.1 Ref.3
Mutagenesis2871H → Q: 54% decrease in activity. Ref.1 Ref.3
Mutagenesis2871H → W: 44% decrease in activity. Ref.1 Ref.3
Mutagenesis2871H → Y: 57% decrease in activity. Ref.1 Ref.3
Mutagenesis3041Q → F: 50% decrease in activity. Ref.3
Mutagenesis3041Q → W: 3% decrease in activity. Ref.3
Mutagenesis3141F → A: 3% increase in activity. Ref.3

Secondary structure

............................................................... 314
Helix Strand Turn

Details...

Sequences

Sequence LengthMass (Da)Tools
Q7SIG4 [UniParc].

Last modified December 15, 2003. Version 1.
Checksum: 8A3F3D84002A6EA8

FASTA31435,080
        10         20         30         40         50         60 
MEIPVIEPLF TKVTEDIPGA EGPVFDKNGD FYIVAPEVEV NGKPAGEILR IDLKTGKKTV 

        70         80         90        100        110        120 
ICKPEVNGYG GIPAGCQCDR DANQLFVADM RLGLLVVQTD GTFEEIAKKD SEGRRMQGCN 

       130        140        150        160        170        180 
DCAFDYEGNL WITAPAGEVA PADYTRSMQE KFGSIYCFTT DGQMIQVDTA FQFPNGIAVR 

       190        200        210        220        230        240 
HMNDGRPYQL IVAETPTKKL WSYDIKGPAK IENKKVWGHI PGTHEGGADG MDFDEDNNLL 

       250        260        270        280        290        300 
VANWGSSHIE VFGPDGGQPK MRIRCPFEKP SNLHFKPQTK TIFVTEHENN AVWKFEWQRN 

       310 
GKKQYCETLK FGIF 

« Hide

References

[1]"Insights into the reaction mechanism of the diisopropyl fluorophosphatase from Loligo vulgaris by means of kinetic studies, chemical modification and site-directed mutagenesis."
Hartleib J., Rueterjans H.
Biochim. Biophys. Acta 1546:312-324(2001) [PubMed] [Europe PMC] [Abstract]
Cited for: BIOPHYSICOCHEMICAL PROPERTIES, MUTAGENESIS OF HIS-181; HIS-219; HIS-224; HIS-248; HIS-274 AND HIS-287.
[2]"Role of calcium ions in the structure and function of the di-isopropylfluorophosphatase from Loligo vulgaris."
Hartleib J., Geschwindner S., Scharff E.I., Rueterjans H.
Biochem. J. 353:579-589(2001) [PubMed] [Europe PMC] [Abstract]
Cited for: CATALYTIC ACTIVITY, COFACTOR, ENZYME REGULATION.
[3]"Mutational and structural studies of the diisopropylfluorophosphatase from Loligo vulgaris shed new light on the catalytic mechanism of the enzyme."
Katsemi V., Luecke C., Koepke J., Lohr F., Maurer S., Fritzsch G., Rueterjans H.
Biochemistry 44:9022-9033(2005) [PubMed] [Europe PMC] [Abstract]
Cited for: FUNCTION, MUTAGENESIS OF GLN-77; ASN-120; ASP-121; TYR-144; ARG-146; MET-148; PHE-173; ASN-175; HIS-181; THR-195; ASP-229; ASP-232; ASN-237; SER-271; ASN-272; HIS-274; HIS-287; GLN-304 AND PHE-314.
[4]"Crystallization and preliminary X-ray crystallographic analysis of DFPase from Loligo vulgaris."
Scharff E.I., Lucke C., Fritzsch G., Koepke J., Hartleib J., Dierl S., Rueterjans H.
Acta Crystallogr. D 57:148-149(2001) [PubMed] [Europe PMC] [Abstract]
Cited for: CRYSTALLIZATION, X-RAY CRYSTALLOGRAPHY (1.8 ANGSTROMS), BIOTECHNOLOGY.
[5]"Crystal structure of diisopropylfluorophosphatase from Loligo vulgaris."
Scharff E.I., Koepke J., Fritzsch G., Lucke C., Rueterjans H.
Structure 9:493-502(2001) [PubMed] [Europe PMC] [Abstract]
Cited for: X-RAY CRYSTALLOGRAPHY (1.8 ANGSTROMS), COFACTOR, SUBUNIT, MUTAGENESIS OF GLU-21; GLU-37; ASP-229; TRP-244 AND SER-271.
[6]"Statistical analysis of crystallographic data obtained from squid ganglion DFPase at 0.85 A resolution."
Koepke J., Scharff E.I., Lucke C., Rueterjans H., Fritzsch G.
Acta Crystallogr. D 59:1744-1754(2003) [PubMed] [Europe PMC] [Abstract]
Cited for: X-RAY CRYSTALLOGRAPHY (0.85 ANGSTROMS), COFACTOR, SUBUNIT.
+Additional computationally mapped references.

Cross-references

3D structure databases

PDBe
RCSB-PDB
PDBj
EntryMethodResolution (Å)ChainPositionsPDBsum
1E1AX-ray1.80A1-314[»]
1PJXX-ray0.85A1-314[»]
2GVUX-ray2.00A1-314[»]
2GVVX-ray1.73A1-314[»]
2GVWX-ray1.86A1-314[»]
2GVXX-ray2.00A1-314[»]
2IAOX-ray2.00A3-314[»]
2IAPX-ray1.90A3-314[»]
2IAQX-ray2.10A3-314[»]
2IARX-ray1.90A3-314[»]
2IASX-ray2.00A3-314[»]
2IATX-ray1.90A3-314[»]
2IAUX-ray2.00A3-314[»]
2IAVX-ray1.07A3-314[»]
2IAWX-ray1.74A3-314[»]
2IAXX-ray1.10A3-314[»]
3BYCX-ray2.20A1-314[»]
3HLHX-ray1.80A/B/C/D1-314[»]
3HLIX-ray1.40A/B/C/D1-314[»]
3I1CX-ray2.20A1-314[»]
3KGGX-ray2.10A1-314[»]
3LI3X-ray1.66A1-314[»]
3LI4X-ray1.35A1-314[»]
3LI5X-ray1.36A1-314[»]
3O4PX-ray0.85A1-314[»]
3U0SX-ray2.60A/B1-314[»]
ProteinModelPortalQ7SIG4.
SMRQ7SIG4. Positions 1-314.
ModBaseSearch...
MobiDBSearch...

Protocols and materials databases

StructuralBiologyKnowledgebaseSearch...

Enzyme and pathway databases

BRENDA3.1.8.2. 3066.
SABIO-RKQ7SIG4.

Family and domain databases

Gene3D2.120.10.30. 1 hit.
InterProIPR011042. 6-blade_b-propeller_TolB-like.
IPR013658. SGL.
[Graphical view]
PfamPF08450. SGL. 1 hit.
[Graphical view]
ProtoNetSearch...

Other

EvolutionaryTraceQ7SIG4.

Entry information

Entry nameDFPA_LOLVU
AccessionPrimary (citable) accession number: Q7SIG4
Entry history
Integrated into UniProtKB/Swiss-Prot: September 5, 2006
Last sequence update: December 15, 2003
Last modified: July 9, 2014
This is version 50 of the entry and version 1 of the sequence. [Complete history]
Entry statusReviewed (UniProtKB/Swiss-Prot)

Relevant documents

PDB cross-references

Index of Protein Data Bank (PDB) cross-references