RGD Reference Report - Identification of paraoxonase 3 in rat liver microsomes: purification and biochemical properties. - Rat Genome Database

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Identification of paraoxonase 3 in rat liver microsomes: purification and biochemical properties.

Authors: Rodrigo, L  Gil, F  Hernandez, AF  Lopez, O  Pla, A 
Citation: Rodrigo L, etal., Biochem J 2003 Nov 15;376(Pt 1):261-8.
RGD ID: 1303961
Pubmed: PMID:12946270   (View Abstract at PubMed)
PMCID: PMC1223763   (View Article at PubMed Central)
DOI: DOI:10.1042/BJ20030732   (Journal Full-text)

Three paraoxonase genes (PON1, PON2 and PON3) have been described so far in mammals. Although considerable information is available regarding PON1, little is known about PON2 and PON3. PON3 has been isolated recently from rabbit serum [Draganov, Stetson, Watson, Billecke and La Du (2000) J. Biol. Chem. 275, 33435-33442] and liver [Ozols (1999) Biochem. J. 338, 265-275]. In the present study, we have identified the presence of PON3 in rat liver microsomes and a method for the purification to homogeneity is presented. PON3 has been purified 177-fold to apparent homogeneity with a final specific activity of 461 units/mg using a method consisting of seven steps: solubilization of the microsomal fraction, hydroxyapatite adsorption, chromatography on DEAE-Sepharose CL-6B, non-specific affinity chromatography on Cibacron Blue 3GA, two DEAE-cellulose steps and a final affinity chromatography on concanavalin A-Sepharose. SDS/PAGE of the final preparation indicated a single protein-staining band with an apparent molecular mass of 43 kDa. The isolated protein was identified by nanoelectrospray MS. Internal amino acid sequences of several peptides were determined and compared with those of human, rabbit and mouse PON3, showing a high similarity. Some biochemical properties of PON3 were also studied, including optimum pH, K(m) and heat and pH stability.

Gene Ontology Annotations    Click to see Annotation Detail View

Biological Process
TermQualifierEvidenceWithReferenceNotesSourceOriginal Reference(s)
coumarin catabolic process  IDA 1303961 RGD 
phenylacetate catabolic process  IDA 1303961 RGD 

Molecular Function
TermQualifierEvidenceWithReferenceNotesSourceOriginal Reference(s)
3,4-dihydrocoumarin hydrolase activity  IDA 1303961 RGD 
arylesterase activity  IDA 1303961substrate: phenylacetate not paraoxonRGD 

Objects Annotated

Genes (Rattus norvegicus)
Pon3  (paraoxonase 3)


Additional Information