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Evaluation of Lipid Droplet Size and Fusion in Bovine Hepatic Cells
Published on: March 10, 2023
Binding of 18-carbon unsaturated fatty acids to bovine β-lactoglobulin--structural and thermodynamic studies
Joanna I Loch1, Piotr Bonarek, Agnieszka Polit
1Faculty of Chemistry, Jagiellonian University, Ingardena 3, 30-060 Kraków, Poland.
Abstract:
Binding of 18-carbon unsaturated oleic and linoleic acid to lactoglobulin, the milk protein, has been studied for the first time by isothermal titration calorimetry (ITC) and X-ray crystallography. Crystal structures determined to resolution 2.10 Å have revealed presence of single fatty acid molecule bound in β-barrel, the primary binding site, with carboxyl group hydrogen bonded to Glu62. The aliphatic chain of both ligands is in almost linear conformation and their interactions with the protein are similar to observed in structure of lactoglobulin with stearic acid. The ITC experiments showed that binding of unsaturated fatty acids to LGB is spontaneous and exothermic. The stoichiometry of binding is lower than 1.0, association constant is 9.7 × 10(5)M(-1) and 9.0 × 10(5)M(-1) for oleic and linoleic acid, respectively. Solvent relief seems to be the major contributor to entropic changes upon fatty acid binding to lactoglobulin.
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