43. Calmodulin regulating calcium sensitivity of Na channels

Abstract

By extrapolating information from existing research and observing previous assumptions regarding the structure of the Na Channel, this experiment was conducted under the hypothesis that the Na Channel is in part regulated by the calmodulin protein, as a result proving calcium sensitivity of the Na Channel. Furthermore, we assume that there is a one to one stoichiometry between the Na Channel and the Calmodulin. There has been extensive research into the functionality and structure of sodium ion channels (Na channels), as several diseases are associated with the lack of regulation of sodium ions, that is caused by the disfunction of these Na channels. However, one highly controversial matter in the field is the importance of the protein calmodulin (CaM) and calcium in Na channel function. Calmodulin is a protein that is well known for its role as a calcium binding messenger protein, and that association is believed to play an indirect role in regulating the Na channel through the Na channel’s supposed calcium sensitivity. While there are proponents for both sides, there has been relatively little research that provides strong evidence for either case. In this experiment, the effect of calmodulin on NaV 1.5 is tested by preparing a set of cardiac cells (of the human specie) with the NaV 1.5 C-Termini and CaM protein, which were then to be placed in solutions with varying concentrations of calcium. We took special care to test multiple concentrations of calcium, as previous studies have tested very low concentrations, with Manu Ben-Johny’s team from the John Hopkins laboratory in particular testing up to a meager 50 micromolar, despite producing a well-respected paper (By comparison, the average Na channel can naturally sustain a concentration of almost 1-2 millimolar and on some occasions, reaching even higher concentrations). After using light scattering and observing the signals given off by the calcium interacting with these Nav1.5/CaM complexes across the varying calcium concentrations, the overall pattern indicated that there was a one to one stoichiometry between calmodulin and Nav 1.5. More importantly, it indicated calcium sensitivity of the Na channel. With this research, a definitive answer has been drawn regarding the importance of calmodulin in calcium modulation in Na channels. Not only does this have the effect of creating a foundation for further research into the structure and function of Na channels, but it also gives deep insight into fundamental functions of the channel that can play a major role into the creation of drugs to treat the many cardiac diseases associated with dysfunction of the channel.