Dynamic control of deactivation gating by a soluble amino-terminal domain in HERG K+ channels

K+ channels encoded by the human ether-a-go-go-related gene (HERG) are dist inguished from most other voltage-gated K+ channels by an unusually slow de activation process that enables cardiac I-KI the corresponding current in v entricular cells, to contribute to the repolarization of the action potenti al. When the first 16 amino acids are deleted from the amino terminus of HE RG, the deactivation rate is much faster (Wang,J., M.C. Trudeau, A.M. Zappi a, and G.A. Robertson. 1998. J. Gen. Physiol. 112:637-647). In this study, we determined whether the first 16 amino acids comprise a functional domain capable of slowing deactivation. We also tested whether this "deactivation subdomain" slows deactivation directly by affecting channel open times or indirectly by a blocking mechanism. Using inside-out macropatches excised f rom Xenopus oocytes, we found that a peptide corresponding to the first 16 amino acids of HERG is sufficient to reconstitute slow deactivation to chan nels lacking the amino terminus. The peptide acts as a soluble domain in a rapid and readily reversible manner, reflecting a more dynamic regulation o f deactivation than the slow modification observed in a previous study with a larger amino-terminal peptide fragment (Morais Cabral, J.H., A. Lee, S.L . Cohen, B.T. Chait, M. Li, and R. Mackinnon. 1998. Cell. 95:649-655). The slowing of deactivation by the peptide occurs in a dose-dependent manner, w ith a Hill coefficient that implies the cooperative action of at least thre e peptides per channel. Unlike internal TEA, which slows deactivation indir ectly by blocking the channels, the peptide does not reduce current amplitu de. Nor does the amino terminus interfere with the blocking effect of TEA, indicating that the amino terminus binding site is spatially distinct from the TEA binding site. Analysis of the single channel activity in cell-attac hed patches shows that the amino terminus significantly increases channel m ean open time with no alteration of the mean closed time or the addition of nonconducting states expected from a pore block mechanism. We propose that the four amino-terminal deactivation subdomains of the tetrameric channel interact with binding sites uncovered by channel opening to specifically st abilize the open state and thus slow channel closing.