Cooperative binding mitigates the high-dose hook effect

• Published in: BMC systems biology Vol. 11; no. 1; p. 74
• Main Authors: Roy, Ranjita Dutta, Rosenmund, Christian, Stefan, Melanie I.
• Format: Journal Article
• Language: English
• Published: London BioMed Central 14.08.2017; BioMed Central Ltd
• Subjects: Algorithms; Allosteric properties; Analysis; Antigens; Binding sites; Bioinformatics; Biomedical and Life Sciences; Ca2+/calmodulin-dependent protein kinase II; Calcium; Calcium - metabolism; Calcium-binding protein; Calmodulin; Calmodulin - metabolism; Cellular and Medical Topics; Combinatorial analysis; Computational Biology/Bioinformatics; Computer simulation; Cooperativity; Expected values; Genetic aspects; Kinases; Kinetics; Life Sciences; Ligands; Long-term depression; Long-term potentiation; Models, Biological; Neurons; pharmacology and medicine; Physiological; Physiological aspects; Protein Binding; Proteins; Research Article; Simulation; Simulation and Modeling; Systems Biology; Systems physiology; United States; Sweden; Germany; Mechanistic model; Prozone effect; Allostery; Cooperativity; High-dose Hook effect; Calmodulin
• ISSN: 1752-0509; 1752-0509
• DOI: 10.1186/s12918-017-0447-8

Background The high-dose hook effect (also called prozone effect) refers to the observation that if a multivalent protein acts as a linker between two parts of a protein complex, then increasing the amount of linker protein in the mixture does not always increase the amount of fully formed complex. On the contrary, at a high enough concentration range the amount of fully formed complex actually decreases. It has been observed that allosterically regulated proteins seem less susceptible to this effect. The aim of this study was two-fold: First, to investigate the mathematical basis of how allostery mitigates the prozone effect. And second, to explore the consequences of allostery and the high-dose hook effect using the example of calmodulin, a calcium-sensing protein that regulates the switch between long-term potentiation and long-term depression in neurons. Results We use a combinatorial model of a “perfect linker protein” (with infinite binding affinity) to mathematically describe the hook effect and its behaviour under allosteric conditions. We show that allosteric regulation does indeed mitigate the high-dose hook effect. We then turn to calmodulin as a real-life example of an allosteric protein. Using kinetic simulations, we show that calmodulin is indeed subject to a hook effect. We also show that this effect is stronger in the presence of the allosteric activator Ca 2+ /calmodulin-dependent kinase II (CaMKII), because it reduces the overall cooperativity of the calcium-calmodulin system. It follows that, surprisingly, there are conditions where increased amounts of allosteric activator actually decrease the activity of a protein. Conclusions We show that cooperative binding can indeed act as a protective mechanism against the hook effect. This will have implications in vivo where the extent of cooperativity of a protein can be modulated, for instance, by allosteric activators or inhibitors. This can result in counterintuitive effects of decreased activity with increased concentrations of both the allosteric protein itself and its allosteric activators.