Bonarek Piotr, Mularczyk Dorota, Loch Joanna, Dziedzicka-Wasylewska Marta
Department of Physical Biochemistry, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Kraków, Poland.
Department of Physical Biochemistry, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Kraków, Poland.
Biophys Chem. 2022 Apr;283:106770. doi: 10.1016/j.bpc.2022.106770. Epub 2022 Jan 31.
β-Lactoglobulin (BLG), a member of the lipocalin family, is a well-studied model protein. It is also widely used as a scaffold for the development of novel proteins. Our previous work adopted a rational approach based on homolog structure alignment to obtain several BLG variants with point mutations inside the binding pocket. To investigate the effect of mutation on ligand binding thermodynamics, we chose a set of aliphatic ligands and performed a study based on isothermal titration calorimetry. In addition, the circular dichroism spectra observed for the protein-ligand complexes were analyzed. The ligand binding thermodynamics was compared between wild-type and mutated BLG as well as between two ligands. The findings pointed to factors that can be responsible for the mutation-induced changes in the thermodynamics of the complexes.
β-乳球蛋白（BLG）是脂质运载蛋白家族的成员，是一种经过充分研究的模型蛋白。它还被广泛用作新型蛋白质开发的支架。我们之前的工作采用了基于同源结构比对的合理方法，以获得结合口袋内有单点突变的几种BLG变体。为了研究突变对配体结合热力学的影响，我们选择了一组脂肪族配体，并基于等温滴定量热法进行了研究。此外，还分析了蛋白质-配体复合物的圆二色光谱。比较了野生型和突变型BLG之间以及两种配体之间的配体结合热力学。研究结果指出了可能导致复合物热力学发生突变诱导变化的因素。
