Mantle redox state drives outgassing chemistry and atmospheric composition of rocky planets.

Volcanic degassing of planetary interiors has important implications for their corresponding atmospheres. The oxidation state of rocky interiors affects the volatile partitioning during mantle melting and subsequent volatile speciation near the surface. Here we show that the mantle redox state is central to the chemical composition of atmospheres while factors such as planetary mass, thermal state, and age mainly affect the degassing rate. We further demonstrate that mantle oxygen fugacity has an effect on atmospheric thickness and that volcanic degassing is most efficient for planets between 2 and 4 Earth masses. We show that outgassing of reduced systems is dominated by strongly reduced gases such as [Formula: see text], with only smaller fractions of moderately reduced/oxidised gases ([Formula: see text], [Formula: see text]). Overall, a reducing scenario leads to a lower atmospheric pressure at the surface and to a larger atmospheric thickness compared to an oxidised system. Atmosphere predictions based on interior redox scenarios can be compared to observations of atmospheres of rocky exoplanets, potentially broadening our knowledge on the diversity of exoplanetary redox states.

Optical observations of comet Hale-Bopp (C/1995 O1) at large heliocentric distances before perihelion.

The activity of comet Hale-Bopp (C/1995 O1) was monitored monthly by optical imaging and long-slit spectroscopy of its dust and gas distribution over heliocentric distances of 4.6 to 2.9 astronomical units. The observed band intensities of the NH2 radical and the H2O+ ion cannot be explained by existing models of fluorescence excitation, warranting a reexamination of the corresponding production rates, at least at large heliocentric distances. Comparing the production rate of the CN radical to its proposed parent, HCN, shows no evidence for the need of a major additional source for CN in Hale-Bopp at large heliocentric distances. The dust and CN production rates are consistent with a significant amount of sublimation occurring from icy dust grains surrounding Hale-Bopp.

Evolution of the outgassing of comet Hale-Bopp (C/1995 O1) from radio observations.

Spectra obtained from ground-based radio telescopes show the progressive release of CO, CH3OH, HCN, H2O (from OH), H2S, CS, H2CO, CH3CN, and HNC as comet Hale-Bopp (C/1995 01) approached the sun from 6.9 to 1.4 astronomical units (AU). The more volatile species were relatively more abundant in the coma far from the sun, but there was no direct correlation between overabundance and volatility. Evidence for H2O sublimation from icy grains was seen beyond 3.5 AU from the sun. The change from a CO-driven coma to an H2O-driven coma occurred at about 3 AU. The gas outflow velocity and temperature increased as Hale-Bopp approached the sun.

Searching for Atmospheric Bioindicators in Planets around the Two Nearby Stars, Proxima Centauri and Epsilon Eridani-Test Cases for Retrieval of Atmospheric Gases with Infrared Spectroscopy.

We tested the ability of thermal infrared spectroscopy to retrieve assumed atmospheric compositions for different types of planets orbiting Proxima Centauri and Epsilon Eridani. Six cases are considered, covering a range of atmospheric compositions and some diversity in the bulk composition (rocky, water ocean, hydrogen rich) and the spectral type of the parent star (M and K stars). For some cases, we applied coupled climate chemistry, or climate-only calculations; for other cases, we assumed the atmospheric composition, ground temperature, and surface reflectivity. The IR emission was then calculated from line-by-line radiative transfer models and used to investigate retrieval of input atmospheric species. For the six cases considered, no false positive of the triple bioindicator (H2O, CO2, and O2, in specified conditions) was found. In some cases, results show that the simultaneous acquisition of a visible spectrum would be valuable, for example, when CO2 is very abundant and its 9.4 µm satellite band hides the 9.6 µm O3 band in the IR. In each case, determining the mass appears mandatory to identify the planet's nature and have an idea of surface conditions, which are necessary when testing for the presence of life.

Images from the surface of asteroid Ryugu show rocks similar to carbonaceous chondrite meteorites.

The near-Earth asteroid (162173) Ryugu is a 900-m-diameter dark object expected to contain primordial material from the solar nebula. The Mobile Asteroid Surface Scout (MASCOT) landed on Ryugu's surface on 3 October 2018. We present images from the MASCOT camera (MASCam) taken during the descent and while on the surface. The surface is covered by decimeter- to meter-sized rocks, with no deposits of fine-grained material. Rocks appear either bright, with smooth faces and sharp edges, or dark, with a cauliflower-like, crumbly surface. Close-up images of a rock of the latter type reveal a dark matrix with small, bright, spectrally different inclusions, implying that it did not experience extensive aqueous alteration. The inclusions appear similar to those in carbonaceous chondrite meteorites.

Evolution of Earth-like Planetary Atmospheres around M Dwarf Stars: Assessing the Atmospheres and Biospheres with a Coupled Atmosphere Biogeochemical Model.

Earth-like planets orbiting M dwarfs are prominent targets when searching for life outside the Solar System. We apply our Coupled Atmosphere Biogeochemical model to investigate the coupling between the biosphere, geosphere, and atmosphere in order to gain insight into the atmospheric evolution of Earth-like planets orbiting M dwarfs and to understand the processes affecting biosignatures and climate on such worlds. This is the first study applying an automated chemical pathway analysis quantifying the production and destruction pathways of molecular oxygen (O2) for an Earth-like planet with an Archean O2 concentration orbiting in the habitable zone of the M dwarf star AD Leonis, which we take as a type-case of an active M dwarf. The main production arises in the upper atmosphere from carbon dioxide photolysis followed by catalytic hydrogen oxide radical (HOx) reactions. The strongest destruction does not take place in the troposphere, as was the case in Gebauer et al. ( 2017 ) for an early Earth analog planet around the Sun, but instead in the middle atmosphere where water photolysis is the strongest. Results further suggest that these atmospheres are in absolute terms less destructive for O2 than for early Earth analog planets around the Sun despite higher concentrations of reduced gases such as molecular hydrogen, methane, and carbon monoxide. Hence smaller amounts of net primary productivity are required to oxygenate the atmosphere due to a change in the atmospheric oxidative capacity, driven by the input stellar spectrum resulting in shifts in the intrafamily HOx partitioning. Under the assumption that an atmosphere of an Earth-like planet survived and evolved during the early high-activity phase of an M dwarf to an Archean-type composition, a possible "Great Oxidation Event," analogous to that on Early Earth, would have occurred earlier in time after the atmospheric composition was reached, assuming the same atmospheric O2 sources and sinks as on early Earth. Key Words: Earth-like-Oxygen-M dwarf stars-Atmosphere-Biogeochemistry-Photochemistry-Biosignatures-Earth-like planets. Astrobiology 18, 856-872.

Randomized comparison of the efficacy and safety of ciclesonide and budesonide in adolescents with severe asthma.

BACKGROUND: The aim of the study was to investigate the efficacy and safety of ciclesonide compared with budesonide in adolescents with severe asthma. METHODS: In this randomized, double-blind, double-dummy, parallel-group study, patients aged 12-17 years with severe asthma were treated with budesonide 400 microg once daily (QD) in a 2-week run-in period. At randomization, eligible patients were assigned 2:1 to ciclesonide 320 microg QD (ex-actuator) or budesonide 800 microg QD (metered dose), respectively, in the evening. Forced expiratory volume in 1s (FEV(1)) was the primary variable. Patients recorded asthma symptom score and rescue medication use in diaries. Safety assessments included adverse events (AEs) and 24-h urine cortisol. RESULTS: Four hundred and three patients were randomized. Ciclesonide 320 microg QD and budesonide 800 microg QD significantly increased FEV(1) (least-squares mean: 505 and 536 mL, respectively; both p<0.0001 versus baseline) in the intention-to-treat (ITT) population. Lower limits of the 95% confidence intervals (ITT: -138 mL; per-protocol: -122 mL) were above the non-inferiority limit (-150 mL). Median percentage of days without asthma symptoms and without rescue medication use was 84% with ciclesonide and 85% with budesonide. AEs were unremarkable, with no cases of confirmed candidiasis. Median creatinine-adjusted urine cortisol significantly decreased with budesonide treatment (15.9-13.7 nmol cortisol/mmol creatinine; p=0.0086 versus baseline), but not with ciclesonide (p=0.1125). CONCLUSIONS: Ciclesonide 320 microg QD showed similar efficacy to budesonide 800 microg QD in adolescents with severe asthma. Ciclesonide was well tolerated and, unlike budesonide, had no effect on urine cortisol levels. CLINICAL TRIAL REGISTRATION NUMBER: EudraCT No.: 2004-001233-41.

Evolution of Earth-like Extrasolar Planetary Atmospheres: Assessing the Atmospheres and Biospheres of Early Earth Analog Planets with a Coupled Atmosphere Biogeochemical Model.

Understanding the evolution of Earth and potentially habitable Earth-like worlds is essential to fathom our origin in the Universe. The search for Earth-like planets in the habitable zone and investigation of their atmospheres with climate and photochemical models is a central focus in exoplanetary science. Taking the evolution of Earth as a reference for Earth-like planets, a central scientific goal is to understand what the interactions were between atmosphere, geology, and biology on early Earth. The Great Oxidation Event in Earth's history was certainly caused by their interplay, but the origin and controlling processes of this occurrence are not well understood, the study of which will require interdisciplinary, coupled models. In this work, we present results from our newly developed Coupled Atmosphere Biogeochemistry model in which atmospheric O2 concentrations are fixed to values inferred by geological evidence. Applying a unique tool (Pathway Analysis Program), ours is the first quantitative analysis of catalytic cycles that governed O2 in early Earth's atmosphere near the Great Oxidation Event. Complicated oxidation pathways play a key role in destroying O2, whereas in the upper atmosphere, most O2 is formed abiotically via CO2 photolysis. The O2 bistability found by Goldblatt et al. ( 2006 ) is not observed in our calculations likely due to our detailed CH4 oxidation scheme. We calculate increased CH4 with increasing O2 during the Great Oxidation Event. For a given atmospheric surface flux, different atmospheric states are possible; however, the net primary productivity of the biosphere that produces O2 is unique. Mixing, CH4 fluxes, ocean solubility, and mantle/crust properties strongly affect net primary productivity and surface O2 fluxes. Regarding exoplanets, different "states" of O2 could exist for similar biomass output. Strong geological activity could lead to false negatives for life (since our analysis suggests that reducing gases remove O2 that masks its biosphere over a wide range of conditions). Key Words: Early Earth-Proterozoic-Archean-Oxygen-Atmosphere-Biogeochemistry-Photochemistry-Biosignatures-Earth-like planets. Astrobiology 16, 27-54.

Alkoxypsoralens, novel nonpeptide blockers of Shaker-type K+ channels: synthesis and photoreactivity.

A series of psoralens and structurally related 5,7-disubstituted coumarins was synthesized and investigated for their K+ channel blocking activity as well as for their phototoxicity to Artemia salina and their ability to generate singlet oxygen and to photomodify DNA. After screening the compounds on Ranvier nodes of the toad Xenopus laevis, the affinities of the most promising compounds, which proved to be psoralens bearing alkoxy substituents in the 5-position or alkoxymethyl substituents in the neighboring 4- or 4'-position, to a number of homomeric K+ channels were characterized. All compounds exhibited the highest affinity to Kv1.2. 5,8-Diethoxypsoralen (10d) was found to be an equally potent inhibitor of Kv1.2 and Kv1.3, while lacking the phototoxicity normally inherent in psoralens. The reported compounds represent a novel series of nonpeptide blockers of Shaker-type K+ channels that could be further developed into selective inhibitors of Kv1.2 or Kv1. 3.

Chemistry and pharmacology of the non-benzodiazepine anxiolytic enciprazine and related compounds.

In the course of studies on tranquilizers, new non-benzodiazepine-like compounds were synthesized. These are 1-(3,4,5-trimethoxyphenoxy)-3-[4-(2-methoxyphenyl)piperazinyl]prop an-2-ol (INN: enciprazine) and derivatives thereof which were screened pharmacologically in order to evaluate their central nervous system activity. Compounds with marked antiaggressive and anxiolytic properties but without dependence potential could be detected. Enciprazine was selected for clinical investigations.

The effect of deep pore mutations on the action of phenylalkylamines on the Kv1.3 potassium channel.

We investigated the action of the phenylalkylamines verapamil and N-methyl-verapamil on the Kv1.3 potassium channel using the whole-cell configuration of the patch-clamp technique. Our goal was to identify their binding as a prerequisite for using the phenylalkylamines as small, well-defined molecular probes, not only to expand the structural findings made with peptide toxins or by crystallization, but also to use them as lead compounds for the generation of more potent and therefore more specific K+ channel modulators. Competition experiments with charybdotoxin, known to interact with external residues of Kv1.3, showed no interaction with verapamil. The internal application of quarternary N-methyl-verapamil in combination with verapamil suggested competition for the same internal binding site. Verapamil affinity was decreased 6 fold by a mutation (M395V) in a region of the internal pore which forms part of the internal tetraethylammonium (TEA+) binding site, although mutations at neighbouring residues (T396 and T397) were without effect. Modification of C-type inactivation by mutations in the internal pore suggest that this region participates in the inactivation process. The action of phenylalkylamines and local anaesthetics on L-type Ca2+ channels and Na channels, respectively, and verapamil on Kv1.3 indicate very similar blocking mechanisms. This might allow the use of these compounds as molecular probes to map the internal vestibule of all three channel types.

UK-78,282, a novel piperidine compound that potently blocks the Kv1.3 voltage-gated potassium channel and inhibits human T cell activation.

1. UK-78,282, a novel piperidine blocker of the T lymphocyte voltage-gated K+ channel, Kv1.3, was discovered by screening a large compound file using a high-throughput 86Rb efflux assay. This compound blocks Kv1.3 with a IC50 of approximately 200 nM and 1:1 stoichiometry. A closely related compound, CP-190,325, containing a benzyl moiety in place of the benzhydryl in UK-78,282, is significantly less potent. 2 Three lines of evidence indicate that UK-78,282 inhibits Kv1.3 in a use-dependent manner by preferentially blocking and binding to the C-type inactivated state of the channel. Increasing the fraction of inactivated channels by holding the membrane potential at - 50 mV enhances the channel's sensitivity to UK-78,282. Decreasing the number of inactivated channels by exposure to approximately 160 mM external K+ decreases the sensitivity to UK-78,282. Mutations that alter the rate of C-type inactivation also change the channel's sensitivity to UK-78,282 and there is a direct correlation between tau(h) and IC50 values. 3. Competition experiments suggest that UK-78,282 binds to residues at the inner surface of the channel overlapping the site of action of verapamil. Internal tetraethylammonium and external charybdotoxin do not compete UK-78,282's action on the channel. 4. UK-78,282 displays marked selectivity for Kv1.3 over several other closely related K+ channels, the only exception being the rapidly inactivating voltage-gated K+ channel, Kv1.4. 5. UK-78,282 effectively suppresses human T-lymphocyte activation.

The dependence of the ice-albedo feedback on atmospheric properties.

Ice-albedo feedback is a potentially important destabilizing effect for the climate of terrestrial planets. It is based on the positive feedback between decreasing surface temperatures, an increase of snow and ice cover, and an associated increase in planetary albedo, which then further decreases surface temperature. A recent study shows that for M stars, the strength of the ice-albedo feedback is reduced due to the strong spectral dependence of stellar radiation and snow/ice albedos; that is, M stars primarily emit in the near IR, where the snow and ice albedo is low, and less in the visible, where the snow/ice albedo is high. This study investigates the influence of the atmosphere (in terms of surface pressure and atmospheric composition) on this feedback, since an atmosphere was neglected in previous studies. A plane-parallel radiative transfer model was used for the calculation of planetary albedos. We varied CO2 partial pressures as well as the H2O, CH4, and O3 content in the atmosphere for planets orbiting Sun-like and M type stars. Results suggest that, for planets around M stars, the ice-albedo effect is significantly reduced, compared to planets around Sun-like stars. Including the effects of an atmosphere further suppresses the sensitivity to the ice-albedo effect. Atmospheric key properties such as surface pressure, but also the abundance of radiative trace gases, can considerably change the strength of the ice-albedo feedback. For dense CO2 atmospheres of the order of a few to tens of bar, atmospheric rather than surface properties begin to dominate the planetary radiation budget. At high CO2 pressures, the ice-albedo feedback is strongly reduced for planets around M stars. The presence of trace amounts of H2O and CH4 in the atmosphere also weakens the ice-albedo effect for both stellar types considered. For planets around Sun-like stars, O3 could also lead to a very strong decrease of the ice-albedo feedback at high CO2 pressures.

Potential biosignatures in super-Earth atmospheres II. Photochemical responses.

Spectral characterization of super-Earth atmospheres for planets orbiting in the habitable zone of M dwarf stars is a key focus in exoplanet science. A central challenge is to understand and predict the expected spectral signals of atmospheric biosignatures (species associated with life). Our work applies a global-mean radiative-convective-photochemical column model assuming a planet with an Earth-like biomass and planetary development. We investigated planets with gravities of 1g and 3g and a surface pressure of 1 bar around central stars with spectral classes from M0 to M7. The spectral signals of the calculated planetary scenarios have been presented by in an earlier work by Rauer and colleagues. The main motivation of the present work is to perform a deeper analysis of the chemical processes in the planetary atmospheres. We apply a diagnostic tool, the Pathway Analysis Program, to shed light on the photochemical pathways that form and destroy biosignature species. Ozone is a potential biosignature for complex life. An important result of our analysis is a shift in the ozone photochemistry from mainly Chapman production (which dominates in Earth's stratosphere) to smog-dominated ozone production for planets in the habitable zone of cooler (M5-M7)-class dwarf stars. This result is associated with a lower energy flux in the UVB wavelength range from the central star, hence slower planetary atmospheric photolysis of molecular oxygen, which slows the Chapman ozone production. This is important for future atmospheric characterization missions because it provides an indication of different chemical environments that can lead to very different responses of ozone, for example, cosmic rays. Nitrous oxide, a biosignature for simple bacterial life, is favored for low stratospheric UV conditions, that is, on planets orbiting cooler stars. Transport of this species from its surface source to the stratosphere where it is destroyed can also be a key process. Comparing 1g with 3g scenarios, our analysis suggests it is important to include the effects of interactive chemistry.

Evidence for an internal phenylalkylamine action on the voltage-gated potassium channel Kv1.3.

We characterized the action of verapamil and N-methyl-verapamil on current through the delayed-rectifier potassium channel Kv1.3 mouse (mKv1.3). The whole-cell and inside-out configuration of the patch-clamp technique was used to examine the channel properties after injection of in vitro transcribed cRNA into rat basophilic leukemia cells. The action of verapamil on current through mKv1.3 channels could be separated into an acceleration of the rate of current decay during depolarizing pulses and a reduction of steady state peak current when applied either extracellularly or intracellularly. Both effects were greatly reduced when the membrane-impermeable N-methyl-verapamil was applied extracellularly, but it affected current through mKv1.3 channels similar to verapamil if applied to the intracellular side of the membrane. Mutations in the outer vestibule of the mKv1.3 channel did not change the ability of verapamil to accelerate the mKv1.3 current decay during depolarizing pulses, whereas the reduction of the steady state peak current by verapamil applied either extracellularly and intracellularly and by N-methyl-verapamil applied intracellularly was decreased approximately 25-fold in all three cases. Substances known to interact with an extracellular site of the channel (e.g., extracellularly applied tetraethylammonium or kaliotoxin) did not compete with extracellularly applied verapamil on blocking steady state peak current, whereas intracellularly applied tetraethylammonium, which is known to interact with an intracellular site of the channel, was able to reduce the effect of extracellularly applied verapamil on blocking steady state peak current, suggesting competition for a common binding site between verapamil and intracellularly applied tetraethylammonium. The results from the competition experiments as well as from the mutations in the outer vestibule of mKv1.3 are compatible with the idea that verapamil applied extracellularly moves through the membrane to reach its internal binding site on the mKv1.3 channel.

Structural conservation of the pores of calcium-activated and voltage-gated potassium channels determined by a sea anemone toxin.

The structurally defined sea anemone peptide toxins ShK and BgK potently block the intermediate conductance, Ca(2+)-activated potassium channel IKCa1, a well recognized therapeutic target present in erythrocytes, human T-lymphocytes, and the colon. The well characterized voltage-gated Kv1.3 channel in human T-lymphocytes is also blocked by both peptides, although ShK has a approximately 1,000-fold greater affinity for Kv1.3 than IKCa1. To gain insight into the architecture of the toxin receptor in IKCa1, we used alanine-scanning in combination with mutant cycle analyses to map the ShK-IKCa1 interface, and compared it with the ShK-Kv1.3 interaction surface. ShK uses the same five core residues, all clustered around the critical Lys(22), to interact with IKCa1 and Kv1.3, although it relies on a larger number of contacts to stabilize its weaker interactions with IKCa1 than with Kv1.3. The toxin binds to IKCa1 in a region corresponding to the external vestibule of Kv1.3, and the turret and outer pore of the structurally defined bacterial potassium channel, KcsA. Based on the NMR structure of ShK, we deduce the toxin receptor in IKCa1 to have x-y dimensions of approximately 22 A, a diameter of approximately 31 A, and a depth of approximately 8 A; we estimate that the ion selectivity lies approximately 13 A below the outer lip of the toxin receptor. These dimensions are in good agreement with those of the KcsA channel determined from its crystal structure, and the inferred structure of Kv1.3 based on mapping with scorpion toxins. Thus, these distantly related channels exhibit architectural similarities in the outer pore region. This information could facilitate development of specific and potent modulators of the therapeutically important IKCa1 channel.

Nuclear localization and dominant-negative suppression by a mutant SKCa3 N-terminal channel fragment identified in a patient with schizophrenia.

The small conductance calcium-activated K+ channel gene SKCa3/KCNN3 maps to 1q21, a region strongly linked to schizophrenia. Recently, a 4-base pair deletion in SKCa3 was reported in a patient with schizophrenia, which truncates the protein at the end of the N-terminal cytoplasmic region (SKCa3Delta). We generated a green fluorescent protein-SKCa3 N-terminal construct (SKCa3-1/285) that is identical to SKCa3Delta except for the last two residues. Using confocal microscopy we demonstrate that SKCa3-1/285 localizes rapidly and exclusively to the nucleus of mammalian cells like several other pathogenic polyglutamine-containing proteins. This nuclear targeting is mediated in part by two polybasic sequences present at the C-terminal end of SKCa3-1/285. In contrast, full-length SKCa3, SKCa2, and IKCa1 polypeptides are all excluded from the nucleus and express as functional channels. When overexpressed in human Jurkat T cells, SKCa3-1/285 can suppress endogenous SKCa2 currents but not voltage-gated K+ currents. This dominant-negative suppression is most likely mediated through the co-assembly of SKCa3-1/285 with native subunits and the formation of non-functional tetramers. The nuclear localization of SKCa3-1/285 may alter neuronal architecture, and its ability to dominantly suppress endogenous small conductance K(Ca) currents may affect patterns of neuronal firing. Together, these two effects may play a part in the pathogenesis of schizophrenia and other neuropsychiatric disorders.

Regulation of mammalian Shaker-related K+ channels: evidence for non-conducting closed and non-conducting inactivated states.

1. Using the whole-cell recording mode we have characterized two non-conducting states in mammalian Shaker-related voltage-gated K+ channels induced by the removal of extracellular potassium, K+o. 2. In the absence of K+o, current through Kv1.4 was almost completely abolished due to the presence of a charged lysine residue at position 533 at the entrance to the pore. Removal of K+o had a similar effect on current through Kv1.3 when the histidine at the homologous position (H404) was protonated (pH 6.0). Channels containing uncharged residues at the corresponding position (Kv1.1: Y; Kv1.2: V) did not exhibit this behaviour. 3. To characterize the nature of the interaction between Kv1.3 and K+o concentration ([K+]o), we replaced H404 with amino acids of different character, size and charge. Substitution of hydrophobic residues (A, V and L) either in all four subunits or in only two subunits in the tetramer made the channel insensitive to the removal of K+o, possibly by stabilizing the channel complex. Replacement of H404 with the charged residue arginine, or the polar residue asparagine, enhanced the sensitivity of the channel to 0 mM K+o, possibly by making the channel unstable in the absence of K+o. Mutation at a neighbouring position (400) had a similar effect. 4. The effect of removing K+o on current amplitude does not seem to be correlated with the rate of C-type inactivation since the slowly inactivating G380F mutant channel exhibited a similar [K+]o dependence as the wild-type Kv1.3 channel. 5. CP-339,818, a drug that recognizes only the inactivated conformation of Kv1.3, could not block current in the absence of K+o unless the channels were inactivated through depolarizing pulses. 6. We conclude that removal of K+o induces the Kv1.3 channel to transition to a non-conducting 'closed' state which can switch into a non-conducting 'inactivated' state upon depolarization.

Designed peptide analogues of the potassium channel blocker ShK toxin.

ShK toxin, a potassium channel blocker from the sea anemone Stichodactyla helianthus, is a 35-residue polypeptide cross-linked by 3 disulfide bridges. In an effort to generate truncated peptidic analogues of this potent channel blocker, we have evaluated three analogues, one in which the native sequence was truncated and then stabilized by the introduction of additional covalent links (a non-native disulfide and two lactam bridges), and two in which non-native structural scaffolds stabilized by disulfide and/or lactam bridges were modified to include key amino acid residues from the native toxin. The effect of introducing a lactam bridge in the first helix of ShK toxin (to create cyclo14/18[Lys14,Asp18]ShK) was also examined to confirm that this modification was compatible with activity. All four analogues were tested in vitro for their ability to block Kv1.3 potassium channels in Xenopus oocytes, and their solution structures were determined using 1H NMR spectroscopy. The lactam bridge in full-length ShK is well tolerated, with only a 5-fold reduction in binding to Kv1.3. The truncated and stabilized analogue was inactive, apparently due to a combination of slight deviations from the native structure and alterations to side chains required for binding. One of the peptide scaffolds was also inactive because it failed to adopt the required structure, but the other had a K(d) of 92 microM. This active peptide incorporated mimics of Lys22 and Tyr23, which are essential for activity in ShK, and an Arg residue that could mimic Arg11 or Arg24 in the native toxin. Modification of this peptide should produce a more potent, low molecular weight peptidic analogue which will be useful not only for further in vitro and in vivo studies of the effect of blocking Kv1.3, but also for mapping the interactions with the pore and vestibule of this K(+) channel that are required for potent blockade.