Παρασκευή 4 Μαρτίου 2016

Determination of the binding properties of the uremic toxin phenylacetic acid to Human Serum Albumin

Publication date: Available online 3 March 2016
Source:Biochimie
Author(s): Juliana F. Saldanha, Dan Yi, Milena B. Stockler-Pinto, Hédi A. Soula, Stéphane Chambert, Denis Fouque, Denise Mafra, Christophe O. Soulage
Uremic toxins are compounds normally excreted in urine that accumulate in patients with chronic kidney disease as a result of decreased renal clearance. Phenylacetic acid (PAA) has been identified as a new protein bound uremic toxin. The purpose of this study was to investigate in vitro the interaction between PAA and human serum albumin (HSA) at physiological and pathological concentrations. We used ultrafiltration to show that there is a single high-affinity binding site for PAA on HSA, with a binding constant on the order of 3.4×104 M-1 and a maximal stoichiometry of 1.61 mole per mole. The PAA, at the concentration reported in end-stage renal patients, was 26% bound to albumin. Fluorescent probe competition experiments demonstrated that PAA did not bind to Sudlow’s site I (in subdomain IIA) and only weakly bind to Sudlow‘s site II (in subdomain IIIA). The PAA showed no competition with other protein-bound uremic toxins such as p-cresyl-sulfate or indoxyl sulphate for binding to serum albumin. Our results provide evidence that human serum albumin can act as carrier protein for phenylacetic acid.



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