The hydrolysis of 4-nitrophenyl esters of hexanoate (NphOHe) and decanoate (NphODe) by human serum albumin (HSA) at Tyr411, located at the FA3-FA4 site, has been investigated between pH 5.8 and 9.5, at 22.0 degrees C. Values of K-s, k(+2), and k(+2)/K-s obtained at [HSA] >= 5x[NphOXx] and [NphOXx] >= 5x[HSA] (Xx is NphOHe or NphODe) match very well each other; moreover, the deacylation step turns out to be the rate limiting step in catalysis (i.e., k(+3) << k(+2)). The pH dependence of the kinetic parameters for the hydrolysis of NphOHe and NphODe can be described by the acidic pK(a)-shift of a single amino acid residue, which varies from 8.9 in the free HSA to 7.6 and 7.0 in the HSA: NphOHe and HSA: NphODe complex, respectively; the pK>(a)-shift appears to be correlated to the length of the fatty acid tail of the substrate. The inhibition of the HSA-Tyr411-catalyzed hydrolysis of NphOHe, NphODe, and 4-nitrophenyl myristate (NphOMy) by five inhibitors (i.e., diazepam, diflunisal, ibuprofen, 3-indoxyl-sulfate, and propofol) has been investigated at pH 7.5 and 22.0 degrees C, resulting competitive. The affinity of diazepam, diflunisal, ibuprofen, 3-indoxyl-sulfate, and propofol for HSA reflects the selectivity of the FA3-FA4 cleft. Under conditions where Tyr411 is not acylated, the molar fraction of diazepam, diflunisal, ibuprofen, and 3-indoxylsulfate bound to HSA is higher than 0.9 whereas the molar fraction of propofol bound to HSA is ca. 0.5.

Ascenzi, P., Leboffe, L., Masi, A.d., Trezza, V., Fanali, G., Gioia, M., et al. (2015). Ligand binding to the FA3-FA4 cleft inhibits the esterase-like activity of human serum albumin. PLOS ONE, 10(3), e0120603 [10.1371/journal.pone.0120603].

Ligand binding to the FA3-FA4 cleft inhibits the esterase-like activity of human serum albumin

Gioia M.;Coletta M.;
2015-01-01

Abstract

The hydrolysis of 4-nitrophenyl esters of hexanoate (NphOHe) and decanoate (NphODe) by human serum albumin (HSA) at Tyr411, located at the FA3-FA4 site, has been investigated between pH 5.8 and 9.5, at 22.0 degrees C. Values of K-s, k(+2), and k(+2)/K-s obtained at [HSA] >= 5x[NphOXx] and [NphOXx] >= 5x[HSA] (Xx is NphOHe or NphODe) match very well each other; moreover, the deacylation step turns out to be the rate limiting step in catalysis (i.e., k(+3) << k(+2)). The pH dependence of the kinetic parameters for the hydrolysis of NphOHe and NphODe can be described by the acidic pK(a)-shift of a single amino acid residue, which varies from 8.9 in the free HSA to 7.6 and 7.0 in the HSA: NphOHe and HSA: NphODe complex, respectively; the pK>(a)-shift appears to be correlated to the length of the fatty acid tail of the substrate. The inhibition of the HSA-Tyr411-catalyzed hydrolysis of NphOHe, NphODe, and 4-nitrophenyl myristate (NphOMy) by five inhibitors (i.e., diazepam, diflunisal, ibuprofen, 3-indoxyl-sulfate, and propofol) has been investigated at pH 7.5 and 22.0 degrees C, resulting competitive. The affinity of diazepam, diflunisal, ibuprofen, 3-indoxyl-sulfate, and propofol for HSA reflects the selectivity of the FA3-FA4 cleft. Under conditions where Tyr411 is not acylated, the molar fraction of diazepam, diflunisal, ibuprofen, and 3-indoxylsulfate bound to HSA is higher than 0.9 whereas the molar fraction of propofol bound to HSA is ca. 0.5.
2015
Pubblicato
Rilevanza internazionale
Articolo
Esperti anonimi
Settore BIO/10 - BIOCHIMICA
English
Con Impact Factor ISI
Diazepam; Diflunisal; Esterases; Esters; Humans; Hydrogen-Ion Concentration; Hydrolysis; Ibuprofen; Models, Molecular; Propofol; Serum Albumin; Tyrosine
Ascenzi, P., Leboffe, L., Masi, A.d., Trezza, V., Fanali, G., Gioia, M., et al. (2015). Ligand binding to the FA3-FA4 cleft inhibits the esterase-like activity of human serum albumin. PLOS ONE, 10(3), e0120603 [10.1371/journal.pone.0120603].
Ascenzi, P; Leboffe, L; Masi, Ad; Trezza, V; Fanali, G; Gioia, M; Coletta, M; Fasano, M
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/2108/228972
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