An Alternative Purification Method for Human Serum Paraoxonase 1 and its Interactions with Sulfonamides
CHEMICAL BIOLOGY & DRUG DESIGN, cilt.76, sa.6, ss.552-558, 2010 (SCI-Expanded, Scopus)
- Yayın Türü: Makale / Kısa Makale
- Cilt numarası: 76 Sayı: 6
- Basım Tarihi: 2010
- Doi Numarası: 10.1111/j.1747-0285.2010.01036.x
- Dergi Adı: CHEMICAL BIOLOGY & DRUG DESIGN
- Derginin Tarandığı İndeksler: Science Citation Index Expanded (SCI-EXPANDED), Scopus
- Sayfa Sayıları: ss.552-558
- Anahtar Kelimeler: drug target, enzyme purification, inhibition, paraoxonase, sulfonamides, CARBONIC-ANHYDRASE INHIBITORS, LOW-DENSITY-LIPOPROTEIN, IN-VITRO INHIBITION, PROTEASE INHIBITORS, RAT-LIVER, PON1 ACTIVITY, ERYTHROCYTE, VIVO, DRUGS, ARYLESTERASE
- Ondokuz Mayıs Üniversitesi Adresli: Evet
Özet
Paraoxonase 1 (PON1), a high-density lipoprotein (HDL)-associated esterase, is known to mediate antioxidant and antiatherogenic properties. Purification of PON1 has been challenging for a long time. Here, we report a novel purification technique for this enzyme, which allowed us to obtain human serum paraoxonase 1 (hPON1) using straightforward chromatographic methods, such as Diethylaminoethyl-Sephadex anion exchange chromatography and Sepharose 4B-4-phenylazo-2-naphthaleneamine hydrophobic interaction chromatography. We purified the enzyme 302-fold with a final specific activity of 4775 U/mg and a yield of 32%. Furthermore, we examined the in vitro effects of some sulfonamide derivatives, such as sulfacetamide, homosulfanilamide (mafenide), sulfosalazine, furosemide, acetazolamide, and 1,3,4-thiadiazole-2-sulfonamide on the enzyme activity to better understand the inhibitory properties of the molecules. The six sulfonamides dose-dependently decreased the activity of hPON1 with inhibition constants in the millimolar - micromolar range. This study provides an efficient method, which may be useful for other enzymes such as those related to acetylcholinesterase. It also demonstrates the off-target activity of sulfonamides.