An Alternative Purification Method for Human Serum Paraoxonase 1 and its Interactions with Sulfonamides

EKINCI D., SENTURK M., Beydemir S., Kufrevioglu O. I., SUPURAN C. T.

CHEMICAL BIOLOGY & DRUG DESIGN, cilt.76, sa.6, ss.552-558, 2010 (SCI-Expanded) identifier identifier identifier

  • 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
  • Atatürk 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.