Binding study of the fluorescence probe 1-anilino-8-naphthalensulfonate to human plasma and human and bovine serum albumin using potentiometric titration.
The binding of 1-anilino-8-naphthalenesulfonate (ANS) to bovine serum albumin (BSA), human serum albumin (HSA), and human plasma has been studied by potentiometric titration utilizing a laboratory constructed ion selective electrode (ISE) of ANS. Three classes of ANS binding sites were found on BSA, HSA, and plasma at 25 and 37 degrees C. Computer analysis of the data resulted in estimates for the association constants, number of binding sites (HSA, BSA), and binding capacity of each class. The association constants for the first class of binding sites at 25 degrees C were found to be 7.53 (+/- 0.59) x 10(5), 2.70 (+/- 0.20) x 10(5), and 2.64 (+/- 0.26) x 10(5) M-1 for BSA, HSA, and plasma, respectively. Lower values for the association constants of all binding classes were estimated at the higher temperature (37 degrees C). The binding capacity for ANS decreased in the order BSA, plasma, HSA.
Valsami
G N
GN
Department of Pharmacy, University of Athens, Greece.
Macheras
P E
PE
Koupparis
M A
MA
D000817:Anilino Naphthalenesulfonates / Q000097:blood*; D000818:Animals; D002417:Cattle; D004566:Electrodes; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007202:Indicators and Reagents; D011199:Potentiometry; D011485:Protein Binding; D012709:Serum Albumin / Q000378:metabolism*; D012710:Serum Albumin, Bovine / Q000378:metabolism
D000817:Anilino Naphthalenesulfonates; D007202:Indicators and Reagents; D012709:Serum Albumin; D012710:Serum Albumin, Bovine; C027132:1-anilino-8-naphthalenesulfonate