| Popis: |
A critical part of ion channel function is the ability to open and close in response to stimuli, and thus conduct ions in a regulated fashion. While X-ray diffraction studies of ion channels suggested a general steric gating mechanism located at the helix bundle crossing (HBC), recent functional studies on several channels indicate that the helix bundle crossing is open even in closed, non-conductive channels. Two NaK channel variants were crystallized in very different, open and closed conformations and served as an important model of the HBC gating hypothesis. However, neither of these NaK variants are conductive in liposomes unless phenylalanine 92 is mutated to alanine (F92A). Here we use NMR to probe distances at near-atomic resolution of the two NaK variants in lipid bicelles. We demonstrate that in contrast to the crystal structures, both NaK variants are in a fully open conformation, akin to the well known MthK channel structure were the HBC is widely open. Further inquiry into the gating mechanism suggests that the selectivity filter and pore helix are coupled to the M2 helix below and undergo changes in structure when F92 is mutated. Overall, our data shows that NaK exhibits coupling between the selectivity filter and HBC similar to K+ channels and has a more complex gating mechanism than previously thought. |
