Metal Bridge in S4 Segment Supports Helix Transition in Shaker Channel

Carlos A.Z. Bassetto, João Luis Carvalho-de-Souza, Francisco Bezanilla

Research output: Contribution to journalArticlepeer-review

5 Scopus citations


Voltage-gated ion channels play important roles in physiological processes, especially in excitable cells, in which they shape the action potential. In S4-based voltage sensors voltage-gated channels, a common feature is shared; the transmembrane segment 4 (S4) contains positively charged residues intercalated by hydrophobic residues. Although several advances have been made in understating how S4 moves through a hydrophobic plug upon voltage changes, the possible helix transition from α- to 310-helix in S4 during the activation process is still unresolved. Here, we have mutated several hydrophobic residues from I360 to F370 in the S4 segment into histidine, in i, i + 3 and i, i + 6 or i, i + 4 and i, i + 7 pairs, to favor 310- or α-helical conformations, respectively. We have taken advantage of the ability of His to coordinate Zn2+ to promote metal ion bridges, and we have found that the histidine introduced at position 366 (L366H) can interact with the introduced histidine at position 370 (stabilizing that portion of the S4 segment in α-helical conformation). In the presence of 20 μM of Zn2+, the activation currents of L366H:F370H channels were slowed down by a factor of 3.5, and the voltage dependence is shifted by 10 mV toward depolarized potentials with no change on the deactivation time constant. Our data supports that by stabilizing a region of the S4 segment in α-helical conformation, a closed (resting or intermediate) state is stabilized rather than destabilizing the open (active) state. Taken together, our data indicates that S4 undergoes α-helical conformation to a short-lived different secondary structure transiently before reaching the active state in the activation process.

Original languageEnglish (US)
Pages (from-to)922-933
Number of pages12
JournalBiophysical Journal
Issue number4
StatePublished - Feb 25 2020
Externally publishedYes

ASJC Scopus subject areas

  • Biophysics


Dive into the research topics of 'Metal Bridge in S4 Segment Supports Helix Transition in Shaker Channel'. Together they form a unique fingerprint.

Cite this