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Thermal stabilization of human albumin by medium‐ and short‐chain n ‐alkyl fatty acid anions
Author(s) -
Shrake Andrew,
Frazier Douglas,
Schwarz Frederick P.
Publication year - 2006
Publication title -
biopolymers
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.556
H-Index - 125
eISSN - 1097-0282
pISSN - 0006-3525
DOI - 10.1002/bip.20406
Subject(s) - chemistry , propionate , alkyl , formate , denaturation (fissile materials) , differential scanning calorimetry , fatty acid , thermal stability , crystallography , ligand (biochemistry) , organic chemistry , nuclear chemistry , thermodynamics , biochemistry , receptor , physics , catalysis
A comprehensive study of the thermal stabilization of defatted human albumin monomer by n ‐alkyl fatty acid anions (FAAs), formate through n ‐decanoate, was carried out by differential scanning calorimetry (DSC). The concentration of each ligand affording maximum thermal stabilization was determined; n ‐nonanoate provides the greatest stabilization but is only marginally better than n ‐octanoate and n ‐decanoate. The use of reversible thermodynamics and a two‐state denaturation model for albumin has been validated. Standard free energies of binding, calculated from increases in free energy of denaturation, for n ‐butanoate and longer FAAs, are linear with n ‐alkyl chain length whereas those for formate, acetate, and n ‐propionate deviate from linearity; those for acetate and n ‐propionate are even greater than that of n ‐butanoate, thereby suggesting, in addition to the common class of sites available to all such ligands, the presence of an additional class of lower affinity binding sites available only to these shortest ligands. Competition experiments involving acetate and n ‐octanoate and involving n ‐pentanoate and n ‐octanoate confirmed the binding of acetate to lower affinity sites unavailable to n ‐octanoate and n ‐pentanoate. Furthermore, an equation is provided, allowing computation of the transition temperature as a function of the free energy for any reversible process causing a change in thermal stability of a protein undergoing reversible, two‐state denaturation. With this equation, modeling the competition experiments by using the binding parameters determined by DSC provides additional support for the class of lower affinity sites, which play a significant role in thermal stabilization of albumin at higher concentrations of these shortest FAAs. © 2005 Wiley Periodicals, Inc. Biopolymers 81: 235–248, 2006 This article was originally published online as an accepted preprint. The “Published Online” date corresponds to the preprint version. You can request a copy of the preprint by emailing the Biopolymers editorial office at biopolymers@wiley.com