S1 constrains S4 in the voltage sensor domain of Kv7.1 K+ channels

Voltage-gated K(+) channels comprise a central pore enclosed by four voltage-sensing domains (VSDs). While movement of the S4 helix is known to couple to channel gate opening and closing, the nature of S4 motion is unclear. Here, we substituted S4 residues of Kv7.1 channels by cysteine and recorded whole-cell mutant channel currents in Xenopus oocytes using the two-electrode voltage-clamp technique. In the closed state, disulfide and metal bridges constrain residue S225 (S4) nearby C136 (S1) within the same VSD. In the open state, two neighboring I227 (S4) are constrained at proximity while residue R228 (S4) is confined close to C136 (S1) of an adjacent VSD. Structural modeling predicts that in the closed to open transition, an axial rotation (approximately 190 degrees) and outward translation of S4 (approximately 12 A) is accompanied by VSD rocking. This large sensor motion changes the intra-VSD S1-S4 interaction to an inter-VSD S1-S4 interaction. These constraints provide a ground for cooperative subunit interactions and suggest a key role of the S1 segment in steering S4 motion during Kv7.1 gating.     

KCNE1 constrains the voltage sensor of Kv7.1 K+ channels

Kv7 potassium channels whose mutations cause cardiovascular and neurological disorders are members of the superfamily of voltage-gated K(+) channels, comprising a central pore enclosed by four voltage-sensing domains (VSDs) and sharing a homologous S4 sensor sequence. The Kv7.1 pore-forming subunit can interact with various KCNE auxiliary subunits to form K(+) channels with very different gating behaviors. In an attempt to characterize the nature of the promiscuous gating of Kv7.1 channels, we performed a tryptophan-scanning mutagenesis of the S4 sensor and analyzed the mutation-induced perturbations in gating free energy. Perturbing the gating energetics of Kv7.1 bias most of the mutant channels towards the closed state, while fewer mutations stabilize the open state or the inactivated state. In the absence of auxiliary subunits, mutations of specific S4 residues mimic the gating phenotypes produced by co-assembly of Kv7.1 with either KCNE1 or KCNE3. Many S4 perturbations compromise the ability of KCNE1 to properly regulate Kv7.1 channel gating. The tryptophan-induced packing perturbations and cysteine engineering studies in S4 suggest that KCNE1 lodges at the inter-VSD S4-S1 interface between two adjacent subunits, a strategic location to exert its striking action on Kv7.1 gating functions.     

An inactivation gate in the selectivity filter of KCNQ1 potassium channels

Inactivation is an inherent property of most voltage-gated K⁺ channels. While fast N-type inactivation has been analyzed in biophysical and structural details, the mechanisms underlying slow inactivation are yet poorly understood. Here, we characterized a slow inactivation mechanism in various KCNQ1 pore mutants, including L273F, which hinders entry of external Ba²⁺ to its deep site in the pore and traps it by slowing its egress. Kinetic studies, molecular modeling, and dynamics simulations suggest that this slow inactivation involves conformational changes that converge to the outer carbonyl ring of the selectivity filter, where the backbone becomes less flexible. This mechanism involves acceleration of inactivation kinetics and enhancement of Ba²⁺ trapping at elevated external K⁺ concentrations. Hence, KCNQ1 slow inactivation considerably differs from C-type inactivation where vacation of K⁺ from the filter was invoked. We suggest that trapping of K⁺ at s₁ due to filter rigidity and hindrance of the dehydration-resolvation transition underlie the slow inactivation of KCNQ1 pore mutants.     

Organic Acid Exudation by Wild Herbs in Response to Elevated Al Concentrations

In acidic soils, monomeric aluminium (Al³⁺) can reach levelsthat are toxic to plants, thus preventing many species fromgrowing there. Organic acids chelate Al and render it non-toxic.It has been shown that exudation of organic acids by Al-tolerantcrops increases their tolerance to Al. We have extended thisobservation to wild plants by comparing the ability of ten herbsto exude organic acids in response to elevated Al levels. Wehypothesized that exudation of organic acids was related tothe ability of plants to grow on Al-rich soils. Two grasseswere grown in rhizotrons in soils with 41 and 63 µM reactiveAl. Organic acids were sampled from root tips connected to anintact plant-root system.Deschampsia flexuosa (L.) Trin. exudedmore malic acid when grown in the soil with the highest Al content.Five forbs and five grasses were also exposed to three Al levels(0, 25 and 75 µM) in a hydroponic system.Rumex acetosellaL. and Viscaria vulgaris Bernh. increased exudation of oxalicacid and Galium saxatile auct. non L. and Veronica officinalisL. increased exudation of citric acid in response to elevatedAl. The distribution of the forbs in the field as describedby soil pH was negatively related to the amount of organic acidsexuded in response to Al. In contrast, none of the grasses exudedhigher amounts of organic acids with increasing Al concentrationin the hydroponic experiment. Copyright 2001 Annals of BotanyCompany Citrate, malate, aluminium detoxification, rhizotron, hydroponics, rhizosphere, Carex pilulifera,Deschampsia flexuosa , Festuca gigantea, Galium saxatile, Geum urbanum, Holcus mollis,Milium effusum , Rumex acetosella, Veronica officinalis, Viscaria vulgaris