Heme Regulates Allosteric Activation of the Slo1 BK Channel

Large conductance calcium-dependent (Slo1 BK) channels are allosterically activated by membrane depolarization and divalent cations, and possess a rich modulatory repertoire. Recently, intracellular heme has been identified as a potent regulator of Slo1 BK channels (Tang, X.D., R. Xu, M.F. Reynolds, M.L. Garcia, S.H. Heinemann, and T. Hoshi. 2003. Nature. 425:531–535). Here we investigated the mechanism of the regulatory action of heme on heterologously expressed Slo1 BK channels by separating the influences of voltage and divalent cations. In the absence of divalent cations, heme generally decreased ionic currents by shifting the channel's G–V curve toward more depolarized voltages and by rendering the curve less steep. In contrast, gating currents remained largely unaffected by heme. Simulations suggest that a decrease in the strength of allosteric coupling between the voltage sensor and the activation gate and a concomitant stabilization of the open state account for the essential features of the heme action in the absence of divalent ions. At saturating levels of divalent cations, heme remained similarly effective with its influence on the G–V simulated by weakening the coupling of both Ca2+ binding and voltage sensor activation to channel opening. The results thus show that heme dampens the influence of allosteric activators on the activation gate of the Slo1 BK channel. To account for these effects, we consider the possibility that heme binding alters the structure of the RCK gating ring and thereby disrupts both Ca2+- and voltage-dependent gating as well as intrinsic stability of the open state

The β1 Subunit Enhances Oxidative Regulation of Large-Conductance Calcium-activated K+ Channels

Oxidative stress may alter the functions of many proteins including the Slo1 large conductance calcium-activated potassium channel (BKCa). Previous results demonstrated that in the virtual absence of Ca2+, the oxidant chloramine-T (Ch-T), without the involvement of cysteine oxidation, increases the open probability and slows the deactivation of BKCa channels formed by human Slo1 (hSlo1) α subunits alone. Because native BKCa channel complexes may include the auxiliary subunit β1, we investigated whether β1 influences the oxidative regulation of hSlo1. Oxidation by Ch-T with β1 present shifted the half-activation voltage much further in the hyperpolarizing direction (−75 mV) as compared with that with α alone (−30 mV). This shift was eliminated in the presence of high [Ca2+]i, but the increase in open probability in the virtual absence of Ca2+ remained significant at physiologically relevant voltages. Furthermore, the slowing of channel deactivation after oxidation was even more dramatic in the presence of β1. Oxidation of cysteine and methionine residues within β1 was not involved in these potentiated effects because expression of mutant β1 subunits lacking cysteine or methionine residues produced results similar to those with wild-type β1. Unlike the results with α alone, oxidation by Ch-T caused a significant acceleration of channel activation only when β1 was present. The β1 M177 mutation disrupted normal channel activation and prevented the Ch-T–induced acceleration of activation. Overall, the functional effects of oxidation of the hSlo1 pore-forming α subunit are greatly amplified by the presence of β1, which leads to the additional increase in channel open probability and the slowing of deactivation. Furthermore, M177 within β1 is a critical structural determinant of channel activation and oxidative sensitivity. Together, the oxidized BKCa channel complex with β1 has a considerable chance of being open within the physiological voltage range even at low [Ca2+]i

Tiam1 interaction with the PAR complex promotes talin-mediated Rac1 activation during polarized cell migration.

Migrating cells acquire front-rear polarity with a leading edge and a trailing tail for directional movement. The Rac exchange factor Tiam1 participates in polarized cell migration with the PAR complex of PAR3, PAR6, and atypical protein kinase C. However, it remains largely unknown how Tiam1 is regulated and contributes to the establishment of polarity in migrating cells. We show here that Tiam1 interacts directly with talin, which binds and activates integrins to mediate their signaling. Tiam1 accumulated at adhesions in a manner dependent on talin and the PAR complex. The interactions of talin with Tiam1 and the PAR complex were required for adhesion-induced Rac1 activation, cell spreading, and migration toward integrin substrates. Furthermore, Tiam1 acted with talin to regulate adhesion turnover. Thus, we propose that Tiam1, with the PAR complex, binds to integrins through talin and, together with the PAR complex, thereby regulates Rac1 activity and adhesion turnover for polarized migration

Diffractive Interaction and Scaling Violation in pp->pi^0 Interaction and GeV Excess in Galactic Diffuse Gamma-Ray Spectrum of EGRET

We present here a new calculation of the gamma-ray spectrum from pp->pi^0 in the Galactic ridge environment. The calculation includes the diffractive pp interaction and incorporates the Feynman scaling violation for the first time. Galactic diffuse gamma-rays come, predominantly, from pi^0->gamma gamma in the sub-GeV to multi-GeV range. Hunter et al. found, however, an excess in the GeV range ("GeV Excess") in the EGRET Galactic diffuse spectrum above the prediction based on experimental pp->pi^0 cross-sections and the Feynman scaling hypothesis. We show, in this work, that the diffractive process makes the gamma-ray spectrum harder than the incident proton spectrum by ~0.05 in power-law index, and, that the scaling violation produces 30-80% more pi^0 than the scaling model for incident proton energies above 100GeV. Combination of the two can explain about a half of the "GeV Excess" with the local cosmic proton (power-law index ~2.7). The excess can be fully explained if the proton spectral index in the Galactic ridge is a little harder (~0.2 in power-law index) than the local spectrum. Given also in the paper is that the diffractive process enhances e^+ over e^- and the scaling violation gives 50-100% higher p-bar yield than without the violation, both in the multi-GeV range.Comment: 35 pages, 11 figures, to appear in Astrophysical Journa

Impact of intracellular hemin on N-type inactivation of voltage-gated K+ channels

N-type inactivation of voltage-gated K+ channels is conferred by the N-terminal “ball” domains of select pore-forming α subunits or of auxiliary β subunits, and influences electrical cellular excitability. Here, we show that hemin impairs inactivation of K+ channels formed by Kv3.4 α subunits as well as that induced by the subunits Kvβ1.1, Kvβ1.2, and Kvβ3.1 when coexpressed with α subunits of the Kv1 subfamily. In Kvβ1.1, hemin interacts with cysteine and histidine residues in the N terminus (C7 and H10) with high affinity (EC50 100 nM). Similarly, rapid inactivation of Kv4.2 channels induced by the dipeptidyl peptidase-like protein DPP6a is also sensitive to hemin, and the DPP6a mutation C13S eliminates this dependence. The results suggest a common mechanismfor a dynamic regulation of Kv channel inactivation by heme/hemin in N-terminal ball domains of Kv α and auxiliary β subunits. Free intracellular heme therefore has the potential to regulate cellular excitability via modulation of Kv channel inactivation

Evolution of superconductivity in isovalent Te-substituted KxFe2-ySe2 crystals

We report the evolution of superconductivity and the phase diagram of the KxFe2-ySe2-zTez (z=0-0.6) crystals grown by a simple one-step synthesis. No structural transition is observed in any crystals, while lattice parameters exhibit a systematic expansion with Te content. The Tc exhibits a gradual decrease with increasing Te content from Tconset = 32.9 K at z = 0 to Tconset = 27.9 K at z = 0.5, followed by a sudden suppression of superconductivity at z = 0.6. Upon approaching a Te concentration of 0.6, the shielding volume fraction decreases and eventually drops to zero. Simultaneously, hump positions in r-T curve shift to lower temperatures. These results suggest that isovalent substitution of Te for Se in KxFe2-ySe2 crystals suppresses the superconductivity in this system.Comment: 10 pages, 1 table, 8 figure