Application | Comment | Organism |
---|---|---|
medicine | human paraoxonase 1 (h-PON1) is a potential candidate for the development of antidote against organophosphate (OP) compounds poisoning in humans. Insufficient organophosphate-hydrolyzing activity of native enzyme affirms the urgent need to develop improved variant(s) having enhanced organophosphate-hydrolyzing activity | Homo sapiens |
Cloned (Comment) | Organism |
---|---|
gene PON1, recombinant expression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) in inclusion bodies, subcloning in Escherichia coli strain DH5alpha | Homo sapiens |
Protein Variants | Comment | Organism |
---|---|---|
H115W/R192K | site-directed mutagenesis, the mutant shows altered substrate specificity compared to wild-type | Homo sapiens |
H115W/R192K/A137T | site-directed mutagenesis, the mutant shows altered substrate specificity compared to wild-type | Homo sapiens |
H115W/R192K/A137T/D94H/S211T | site-directed mutagenesis, the mutant shows altered substrate specificity compared to wild-type | Homo sapiens |
H115W/R192K/A137T/L130F | site-directed mutagenesis, the mutant shows altered substrate specificity compared to wild-type | Homo sapiens |
H115W/R192K/A137T/M127I/D263H | site-directed mutagenesis, the mutant shows altered substrate specificity compared to wild-type | Homo sapiens |
H115W/R192K/A137T/S81R/P165A | site-directed mutagenesis, the mutant shows altered substrate specificity compared to wild-type | Homo sapiens |
L55M | natural polymorphism, the polymorphism at the 55th position of h-PON1 does not affect the catalytic properties of the enzyme | Homo sapiens |
additional information | h-PON1 is a polymorphic enzyme. A random mutagenesis approach is used to increase the organophosphate (OP)-hydrolyzing activity of recombinant enzyme h-PON1. The mutants not only show a 10-340fold increased OP-hydrolyzing activity against different OP substrates but also exhibit differential lactonase and arylesterase activities, molecular docking studies, overview. Random mutagenesis using Escherichia coli XL-1 Red mutator strain. All mutations result in a considerable decrease in the delta-valerolactone-hydrolyzing activity of the enzyme | Homo sapiens |
R192E | natural polymorphism, polymorphism at position 192 plays an important role in determining the substrate specificity and catalytic efficiency of the enzyme | Homo sapiens |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
2-hydroxyquinoline | a specific reversible competitive inhibitor of h-PON1 that is known to bind in the active site of the enzyme and inhibit the hydrolytic activities of the enzyme | Homo sapiens |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Ca2+ | dependent on | Homo sapiens |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
chlorpyrifos oxon + H2O | Homo sapiens | - |
diethyl phosphate + 3,5,6-trichloropyridin-2-ol | - |
? | |
diisopropyl fluorophosphate + H2O | Homo sapiens | - |
diisopropyl phosphate + fluoride | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Homo sapiens | P27169 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant wild-type and mutant enzymes refolded from Escherichia coli strain BL21(DE3) inclusion bodies by ion exchange chromatography | Homo sapiens |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
diisopropyl fluorophosphate + H2O = diisopropyl phosphate + fluoride | molecular details of the catalytic mechanism of h-PON1 | Homo sapiens |
Renatured (Comment) | Organism |
---|---|
refolding of recombinant wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) inclusion bodies. The recombinant proteins are refolded to their active form by in vitro refolding, and the active protein present in the refolding reaction mixture is further purified | Homo sapiens |
Source Tissue | Comment | Organism | Textmining |
---|---|---|---|
serum | - |
Homo sapiens | - |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
chlorpyrifos oxon + H2O | - |
Homo sapiens | diethyl phosphate + 3,5,6-trichloropyridin-2-ol | - |
? | |
chlorpyrifos oxon + H2O | i.e. CPO, a metabolite of chlorpyrifos that is used as a pesticide in agriculture industry | Homo sapiens | diethyl phosphate + 3,5,6-trichloropyridin-2-ol | - |
? | |
diethyl-paraoxon + H2O | reaction of EC 3.1.8.1 | Homo sapiens | diethyl phosphate + 4-nitrophenol | - |
? | |
diisopropyl fluorophosphate + H2O | - |
Homo sapiens | diisopropyl phosphate + fluoride | - |
? | |
diisopropyl fluorophosphate + H2O | highly toxic structural analogue of G-class type of nerve agents | Homo sapiens | diisopropyl phosphate + fluoride | - |
? | |
additional information | the enzyme is also active with substrates of EC 3.1.1.81, quorum-quenching N-acyl-homoserine lactonase, and EC 3.1.8.1, paraoxonase, hydrolyzing organophosphorus compounds. Measurement of N-oxodecanoyl-DL-homoserine lactone (3O-C10AHL)-hydrolyzing activity (EC 3.1.1.81) of recombinant h-PON1 enzymes is determined by using a recombinant quorum-sensing reporter Escherichia coli strain | Homo sapiens | ? | - |
? | |
phenyl acetate + H2O | - |
Homo sapiens | phenol + acetate | - |
? |
Subunits | Comment | Organism |
---|---|---|
? | x * 45000, about, SDS-PAGE | Homo sapiens |
More | features observed in the structure of PON1, i.e. the six-bladed beta-propeller scaffold, the three alpha-helices at the top of the propeller and the putative calcium-binding residues, are well conserved in the modelled structures | Homo sapiens |
Synonyms | Comment | Organism |
---|---|---|
h-PON1 | - |
Homo sapiens |
More | cf. EC 3.1.8.1 and EC 3.1.1.81 | Homo sapiens |
paraoxonase 1 | - |
Homo sapiens |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Homo sapiens |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
8 | - |
assay at | Homo sapiens |
General Information | Comment | Organism |
---|---|---|
malfunction | insufficient organophosphate-hydrolyzing activity of native enzyme affirms the urgent need to develop improved variant(s) having enhanced organophosphate-hydrolyzing activity. Enzyme mutants show altered substrate specificity with increased activity against paraoxon and lactone substrates, overview | Homo sapiens |
additional information | h-PON1 is a polymorphic enzyme. Molecular docking analysis, homology modelling, overview | Homo sapiens |
physiological function | human paraoxonase 1 (h-PON1) is a serum enzyme that can hydrolyze a variety of substrates, including organophosphate (OP) compounds. PON1 can hydrolyze and inactivate a variety of organophosphate (OP) compounds, including certain OP pesticides and nerve agents (NAs). It is a potential candidate for the development of antidote against OP poisoning in humans. The enzyme possesses anti-inflammatory, anti-oxidative, anti-diabetic and quorum sensor-hydrolyzing activities, it is proposed that the lactonase activity of the enzyme is important for these defensive roles, cf. EC 3.1.1.81 | Homo sapiens |