Lysine and the Na(+)/K(+) Selectivity in Mammalian Voltage-Gated Sodium Channels

Voltage-gated sodium (Na(v)) channels are critical in the generation and transmission of neuronal signals in mammals. The crystal structures of several prokaryotic Na(v) channels determined in recent years inspire the mechanistic studies on their selection upon the permeable cations (especially between Na(+) and K(+) ions), a property that is proposed to be mainly determined by residues in the selectivity filter. However, the mechanism of cation selection in mammalian Na(v) channels lacks direct explanation at atomic level due to the difference in amino acid sequences between mammalian and prokaryotic Na(v) homologues, especially at the constriction site where the DEKA motif has been identified to determine the Na(+)/K(+) selectivity in mammalian Na(v) channels but is completely absent in the prokaryotic counterparts. Among the DEKA residues, Lys is of the most importance since its mutation to Arg abolishes the Na(+)/K(+) selectivity. In this work, we modeled the pore domain of mammalian Na(v) channels by mutating the four residues at the constriction site of a prokaryotic Na(v) channel (Na(v)Rh) to DEKA, and then mechanistically investigated the contribution of Lys in cation selection using molecular dynamics simulations. The DERA mutant was generated as a comparison to understand the loss of ion selectivity caused by the K-to-R mutation. Simulations and free energy calculations on the mutants indicate that Lys facilitates Na(+)/K(+) selection by electrostatically repelling the cation to a highly Na(+)-selective location sandwiched by the carboxylate groups of Asp and Glu at the constriction site. In contrast, the electrostatic repulsion is substantially weakened when Lys is mutated to Arg, because of two intrinsic properties of the Arg side chain: the planar geometric design and the sparse charge distribution of the guanidine group.