Volume of Hsp90 Protein-Ligand Binding Determined by Fluorescent Pressure Shift Assay, Densitometry and NMR.
Related Articles Volume of Hsp90 Protein-Ligand Binding Determined by Fluorescent Pressure Shift Assay, Densitometry and NMR.
J Phys Chem B. 2016 Aug 29;
Authors: Toleikis Z, Sirotkin VA, Skvarnavi?ius G, Smirnovien? J, Roumestand C, Matulis D, Petrauskas V
Abstract
Human heat shock protein 90 (Hsp90) is a key player in the homeostasis of the proteome and participates in numerous diseases, such as cancer. For the design of Hsp90 ATP-ase activity inhibitors, it is important to understand the relationship between an inhibitor structure and its inhibition potential. The volume of inhibitor binding is one of the most important such parameters that are rarely being studied. Here, the volumes of several ligand binding to recombinant Hsp90 were obtained by three independent experimental techniques - fluorescent pressure shift assay, vibrating tube densitometry, and high-pressure NMR. Within the error range, all techniques provided similar volumetric parameters of the investigated protein-ligand systems. Protein-ligand binding volumes were negative suggesting that the protein-ligand complex, together with it's hydration shell, occupies less volume than the separate constituents with their hydration shells. Binding volumes of tightly-binding, sub-nanomolar ligands were significantly more negative than weakly-binding, millimolar ligands. The volumes of binding could be useful for the design of inhibitors with desired recognition properties and further development as drugs.
PMID: 27571383 [PubMed - as supplied by publisher]
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