An alternative purification method for human serum paraoxonase 1 and its interactions with sulfonamides.
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.
Atatürk University, Science Faculty, Department of Chemistry, Biochemistry Division, Erzurum, Turkey.
Deniz
D
Ekinci
Murat
M
Sentürk
Sükrü
S
Beydemir
Omar Irfan
OI
Küfrevioğlu
Claudiu T
CT
Supuran
D043303:Aryldialkylphosphatase / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D002623:Chemistry Techniques, Analytical / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D015394:Molecular Structure; D013449:Sulfonamides / Q000737:chemistry* / Q000378:metabolism
D013449:Sulfonamides; D043303:Aryldialkylphosphatase; C468406:PON1 protein, human