BINDING OF SULFAMETHAZINE TO PIG PLASMA-PROTEINS AND ALBUMIN

The interaction of sulphamethazine (SMZ) with pig plasma proteins and albumin was studied by ultrafiltration and equilibrium dialysis, Bindi ng to pig plasma proteins was monophasic (affinity similar to 9.0 mol/ L x 10(3)) and the main binding protein was albumin, At 37 degrees C a nd pH 7.4, the affinity of SMZ for albumin was about 8.0 mol/L x 10(3) and the number of binding sites was estimated as 1.4. Increasing the temperature from 4 to 45 degrees C resulted in a seven-fold decrease i n affinity, and increasing pH from 6.0 to 8.0 enhanced affinity for pi g albumin ten-fold. The free energy of binding (-Delta G) and enthalpy change (-Delta H) were around 5.5 and 5.1 Kcal/mol, respectively. The total entropy change (Delta S) was small and positive, around 2 cal/m ol/degrees K. Studies with the fluorescent probes warfarin and dansyls arcosine, suggest that these bind to separate sites on porcine albumin . SMZ displaced both probes and inhibited the deacetylation of p-nitro phenyl acetate by pig albumin. We conclude that: (1) binding of SMZ to pig plasma proteins and albumin is weak; (2) the interaction with alb umin is exothermic and enthalpy driven, and (3) pig albumin, like othe r mammalian albumins, appears to possess discrete binding sites for wa rfarin and dansylsarcosine, SMZ interacts with both these loci.