Effect of Competing Electron Acceptors on the Reduction of U(VI) by Desulfotomaculum reducens

The biological reduction of soluble U(VI) to the less soluble U(IV) has been proposed as a strategy to remediate uranium-contaminated sites. However, the majority of the contaminated sites contain, in addition to U(VI), competing electron acceptors (CEAs) that can either enhance or inhibit U(VI) reduction. Desulfotomaculum reducens MI-1 is a sulfate-reducing bacterium able to reduce a variety of electron acceptors including U(VI). We characterized U(VI) reduction by D. reducens in the presence of pyruvate and three CEAs: sulfate, nitrate or soluble ferric iron. In the presence of sulfate or ferric iron and U(VI), cell growth was driven by respiration of the CEA. Nitrate was not used as an electron acceptor for growth and vegetative cells grew instead by fermenting pyruvate. Sulfate remaining after sulfate reduction has ceased or the presence of nitrate did not affect U(VI) reduction. However, in the case of sulfate, the addition of H₂ after the depletion of pyruvate greatly enhanced U(VI) reduction. Contrary to sulfate and nitrate, the presence of Fe(II), the product of Fe(III) reduction, abolished U(VI) reduction. The results from this investigation suggest that this microorganism and others with similar characteristics may play a role in U(VI) bioremediation efforts but only after the soluble Fe(II) produced by Fe(III) reduction has been advected away.     

Characterization of Neutrophilic Fe(II)-Oxidizing Bacteria Isolated from the Rhizosphere of Wetland Plants and Description of Ferritrophicum radicicola gen. nov. sp. nov., and Sideroxydans paludicola sp. nov.

Iron deposits (Fe plaque) on wetland plant roots contain abundant microbial populations, including Fe(II)-oxidizing bacteria (FeOB) that have not been cultured previously. In this study, 4 strains of Fe plaque-associated FeOB were isolated from 4 species of wetland plants. All 4 isolates grew in tight association with Fe-oxides, but did not form any identifiable Fe-oxide structures. All strains were obligate lithotrophic Fe(II)-oxidizers that were microaerobic, and were unable to use other inorganic or organic energy sources. One strain, BrT, was shown to fix ¹⁴ CO ₂ at a rate consistent with its requirement for total cell carbon. The doubling times for the strains varied between 9.5 and 15.8 hours. The fatty acid methyl ester (FAME) profiles of 2 strains, BrT and CCJ, revealed that 16:0, 15:1 isoG, and 14:0 were dominant fatty acids. Phylogenetic analysis of the 16S rRNA gene indicated that all the strains were Betaproteobacteria. Two of the strains, BrT and Br-1 belong to a new species, Sideroxydans paludicola; a third strain, LD-1, is related to Sideroxydans lithotrophicus, a recently described species of FeOB. The fourth isolate, Ferritrophicum radicicola, represented a new genus in a new order of Betaproteobacteria, the Ferritrophicales. There are no other cultured isolates in this order. A small subunit rRNA gene-based, cultivation-independent analysis of Typha latifolia collected from a wetland revealed terminal restriction fragment profiles (tRFLP) consistent with the presence of these bacteria in the rhizosphere. These novel organisms likely play an important role in Fe(II) oxidation kinetics and Fe cycling within many terrestrial and freshwater environments.     

高岭土对Fe~(2+)的吸附及其对高岭土中Fe~(3+)生物还原的影响研究

利用异化铁还原微生物可将高岭土中不溶性的Fe3+还原成可溶性的Fe2+,但是此过程中产生的Fe2+能够被高岭土以及异化铁还原微生物吸附,从而影响高岭土中铁的异化还原。本文研究了pH、高岭土量、Fe2+浓度、温度4个因素对高岭土吸附Fe2+的影响;并采用Logistic方程拟合,研究Fe2+及温度对高岭土中Fe3+的生物还原特征。结果表明:pH、高岭土量、浓度、温度4个因素均会影响高岭土吸附Fe2+,当Fe2+吸附在高岭土和微生物菌体表面时,微生物的活性下降,同时高岭土表面Fe3+的生物可利用性也降低,Fe3+生物还原的最大速率减小。     

Competition of Mn2+ and Zn2+ with59Fe2+ and59Fe3+ for the plasma membrane receptors from lactating mouse mammary gland

Nonlabeled MnCl₂ and ZnSO₄ compete with⁵⁹Fe²⁺-ascorbate and⁵⁹Fe ₂ ³⁺ O₃ for transport binding sites situated on the plasma membranes of lactating mouse mammary gland cells. The binding was found to be a process reaching saturation. The heterologous competition used here ruled out the participation of transferrin and to propose that Fe, Mn, and Zn are transported from blood to milk by a mechanism involving one receptor during lactation. Further experiments are necessary to establish the details of the transport mechanism.     

Inhibition of Fe2+- and Fe3+- induced hydroxyl radical production by the iron-chelating drug deferiprone

Deferiprone (L1) is an effective iron-chelating drug that is widely used for the treatment of iron-overload diseases. It is known that in aqueous solutions Fe²⁺ and Fe³⁺ ions can produce hydroxyl radicals via Fenton and photo-Fenton reactions. Although previous studies with Fe²⁺ have reported ferroxidase activity by L1 followed by the formation of Fe³⁺ chelate complexes and potential inhibition of Fenton reaction, no detailed data are available on the molecular antioxidant mechanisms involved. Similarly, in vitro studies have also shown that L1–Fe³⁺ complexes exhibit intense absorption bands up to 800 nm and might be potential sources of phototoxicity. In this study we have applied an EPR spin trapping technique to answer two questions: (1) does L1 inhibit the Fenton reaction catalyzed by Fe²⁺ and Fe³⁺ ions and (2) does UV–Vis irradiation of the L1–Fe³⁺ complex result in the formation of reactive oxygen species. PBN and TMIO spin traps were used for detection of oxygen free radicals, and TEMP was used to trap singlet oxygen if it was formed via energy transfer from L1 in the triplet excited state. It was demonstrated that irradiation of Fe³⁺ aqua complexes by UV and visible light in the presence of spin traps results in the appearance of an EPR signal of the OH spin adduct (TMIO–OH, a(N)=14.15 G, a(H)=16.25 G; PBN–OH, a(N)=16.0 G, a(H)=2.7 G). The presence of L1 completely inhibited the OH radical production. The mechanism of OH spin adduct formation was confirmed by the detection of methyl radicals in the presence of dimethyl sulfoxide. No formation of singlet oxygen was detected under irradiation of L1 or its iron complexes. Furthermore, the interaction of L1 with Fe²⁺ ions completely inhibited hydroxyl radical production in the presence of hydrogen peroxide. These findings confirm an antioxidant targeting potential of L1 in diseases related to oxidative damage.     

Formation of Bacteriochlorophyll Form B820 in Light Harvesting 2 Complexes from Purple Sulfur Bacteria Treated with Dioxane

Treatment of some sulfur bacteria (Allochromatium minutissimum, Thiorhodospira sibirica, and Ectothiorhodospira halovacuolata WN22) with dioxane results in formation of the bacteriochlorophyll form B820 in the light harvesting complex LH2. This form characterized by absorption maximum at 820 nm has the same absorption spectrum as B820 subcomplex from LH1 complex. Appearance of the B820 form was accompanied by a sharp decrease in absorption in the carotenoid region. This phenomenon observed in all LH2 complexes investigated may be attributed to formation of colorless carotenoid aggregates. This is very similar to the previously reported dissociation of the LH1 complex with carotenoids into B820 subcomplexes. Although the B820 form corresponded the bacteriochlorophyll dimer, its circular dichroism spectrum showed that pigment molecules in this dimer exhibit different interaction than those in the B820 subcomplex. The dioxane treatment of LH2 complexes isolated from Rhodopseudomonas palustris bacteria grown under normal or low intensity illumination did not result in formation of such dimers. It is suggested that bacteriochlorophyll B820 formation is related to unique structure of LH2 complexes from the sulfur bacteria.     

Application of an M13 bacteriophage displaying tyrosine on the surface for detection of Fe3+ and Fe2+ ions

Ferric and ferrous ion plays critical roles in bioprocesses,their influences in many fields have not been fully explored due to the lack of methods for quantification of ferric and ferrous ions in biological system or complex matrix.In this study,an M13 bacteriophage(phage) was engineered for use as a sensor for ferric and ferrous ions via the display of a tyrosine residue on the P8 coat protein.The interaction between the specific phenol group of tyrosine and Fe~(3+)./ Fe~(2+).was used as the sensor.Transmission electron microscopy showed aggregation of the tyrosine-displaying phages after incubation with Fe~(3+) and Fe~(2+).The aggregated phages infected the host bacterium inefficiently.This phenomenon could be utilized for detection of ferric and ferrous ions.For ferric ions,a calibration curve ranging from 200 nmol/L to 8 μmol/L with a detection limit of 58 nmol/L was acquired.For ferrous ions,a calibration curve ranging from 800 nmol/L to 8μmol/L with a detection limit of 641.7 nmol/L was acquired.The assay was specific for Fe~((3+)) and Fe~((2+)) when tested against Ni~(2+),Pb~(2+),Zn~(2+),Mn~(2+),Co~(2+),Ca~(2+),Cu~(2+),Cr~(3+),Ba~(2+),and K~+.The tyrosine displaying phage to Fe~(3+) and Fe~(2+) interaction would have plenty of room in application to biomatenals and bionanotechnology.     

Formation of fluorescent carbon nanodots from kitchen wastes and their application for detection of Fe3+

This work reports a scalable synthesis of water‐dispersible fluorescent carbon nanodots based on the simple hydrothermal method (180 °C for 6 h) of kitchen wastes (grape peel for example). We discuss the feasibility of synthesis from kitchen wastes both experimentally and theoretically, and the as‐prepared nanodots have high selectivity for Fe³⁺ ions based on fluorescence quenching which is due to the complexes between nanodots and metal ions. Copyright © 2014 John Wiley & Sons, Ltd.     

微生物驱动硝酸盐还原耦合亚铁氧化成矿过程的锌胁迫

【目的】探究中性厌氧条件下,金属锌影响下硝酸盐依赖型铁氧化菌Pseudomonas stutzeri LS-2驱动的硝酸盐还原耦合亚铁氧化成矿过程机制,对深入理解中性厌氧环境中微生物亚铁氧化驱动的反硝化作用及重金属固定机制具有重要意义。【方法】以不同Zn(Ⅱ)浓度构建LS-2驱动的亚铁氧化成矿体系,分析不同体系中亚铁氧化速率、硝酸盐还原速率以及形成矿物的结构变化规律。【结果】LS-2驱动的硝酸盐还原耦合亚铁氧化成矿过程中,共存Zn(Ⅱ)降低该过程中硝酸盐的还原速率和亚铁氧化速率。同时,随着Zn(Ⅱ)浓度提高,抑制作用增强。微生物亚铁氧化形成的矿物通过吸附、共沉淀和离子置换等过程固定Zn(Ⅱ),降低Zn(Ⅱ)活性。Zn(Ⅱ)浓度对形成的矿物结构有较大的影响:低浓度Zn(Ⅱ)体系中,形成的矿物为纤铁矿;随着Zn(Ⅱ)浓度的提高,矿物结构与结晶度都有一定程度的变化,当Zn(Ⅱ)达到4 mmol/L时,形成的矿物主要为铁锌尖晶石。【结论】明确了重金属锌对LS-2菌株反硝化及亚铁氧化过程的抑制规律,同时阐明了Zn(Ⅱ)浓度对形成矿物结构的影响。研究结果有助于深入认识中性厌氧环境中重金属与微生物驱动的铁循环和反硝化过程的耦合作用,为土壤重金属污染防治提供理论支撑。     

Regulation of InsP3 receptor activity by neuronal Ca2+-binding proteins

Inositol 1,4,5-trisphosphate receptors (InsP(3)Rs) were recently demonstrated to be activated independently of InsP(3) by a family of calmodulin (CaM)-like neuronal Ca(2+)-binding proteins (CaBPs). We investigated the interaction of both naturally occurring long and short CaBP1 isoforms with InsP(3)Rs, and their functional effects on InsP(3)R-evoked Ca(2+) signals. Using several experimental paradigms, including transient expression in COS cells, acute injection of recombinant protein into Xenopus oocytes and (45)Ca(2+) flux from permeabilised COS cells, we demonstrated that CaBPs decrease the sensitivity of InsP(3)-induced Ca(2+) release (IICR). In addition, we found a Ca(2+)-independent interaction between CaBP1 and the NH(2)-terminal 159 amino acids of the type 1 InsP(3)R. This interaction resulted in decreased InsP(3) binding to the receptor reminiscent of that observed for CaM. Unlike CaM, however, CaBPs do not inhibit ryanodine receptors, have a higher affinity for InsP(3)Rs and more potently inhibited IICR. We also show that phosphorylation of CaBP1 at a casein kinase 2 consensus site regulates its inhibition of IICR. Our data suggest that CaBPs are endogenous regulators of InsP(3)Rs tuning the sensitivity of cells to InsP(3).     

Evidence that Fe2+ complexes of 3-aminopicolinate and 3-mercaptopicolinate activate and inhibit phosphoenolpyruvate carboxykinase

3-Mercaptopicolinic acid is known to be an inhibitor of phosphoenolpyruvate carboxykinase and 3-aminopicolinic acid permits Fe²⁺ to activate the enzyme. The potency of mercaptopicolinate is increased by incubating the enzyme with Fe²⁺ prior to assaying for activity. In the present work, the average combining ratio of either pyridine carboxylate with Fe²⁺ at pH 7.5 was determined to be 2:1 when measured by the method of continuous variation of Job or by elemental analysis of the isolated pyridine carboxylate-Fe²⁺ complexes. The ratio of 3-mercaptopicolinate or 3-aminopicolinate to Fe²⁺ that caused the greatest inhibition or activation of purified phosphoenolpyruvate carboxykinase was 2:1. In the absence of Fe²⁺, neither pyridine carboxylate altered the activity of the enzyme. These results indicate that the two pyridine carboxylates can interact with phosphoenolpyruvate carboxykinase as Fe²⁺ coordination complexes.     

Effect of Dy3+ or Eu2+ co‐activator on a BaCa(SO4)2:Ce3+ mixed alkaline earth sulfate phosphor

The individual emission and energy transfer between Ce³⁺ and Eu²⁺ or Dy³⁺ in BaCa(SO₄)₂ mixed alkaline earth sulfate phosphor prepared using a co‐precipitation method is described. The phosphor was characterized by X‐ray diffraction (XRD) and photoluminescence (PL) studies and doped by Ce;Eu and Dy rare earths. All phosphors showed excellent blue–orange emission on excitation with UV light. PL measurements reveal that the emission intensity of Eu²⁺ or Dy³⁺ dopants is greater than when they are co‐doped with Ce³⁺. An efficient Ce³⁺ → Eu²⁺ [²T_2g(4f⁶5d) → ⁸S_7/2(4f₇)] and Ce³⁺ → Dy³⁺ (⁴ F_9/2 → ⁶H_15/2 and ⁴ F_9/2 → ⁶H_13/2) energy transfer takes place in the BaCa(SO₄)₂ host. A strong blue emission peak was observed at 462 nm for Eu²⁺ ions and an orange emission peak at 574 nm for Dy³⁺ ions. Hence, this phosphor may be used as a lamp phosphor. Copyright © 2015 John Wiley & Sons, Ltd.     

Regulation of Neuronal Nitric Oxide and Cyclic GMP Formation by Ca2+

Nitric oxide (NO) acts as a messenger molecule in the CNS by activating soluble guanylyl cyclase. Rat brain synaptosomal NO synthase was stimulated by Ca2+ in a concentration-dependent manner with half-maximal effects observed at 0.3 microM and 0.2 microM when its activity was assayed as formation of NO and L-citrulline, respectively. Cyclic GMP formation was apparently inhibited, however, at Ca2+ concentrations required for the activation of NO synthase, indicating a down-regulation of the signal in NO-producing cells. Purified synaptosomal guanylyl cyclase was not inhibited directly by Ca2+, and the effect was not mediated by a protein binding to guanylyl cyclase at low or high Ca2+ concentrations. In cytosolic fractions, the breakdown of cyclic GMP, but not that of cyclic AMP, was highly stimulated by Ca2+, and 3-isobutyl-1-methylxanthine did not block this reaction effectively. The effects of Ca2+ on cyclic GMP hydrolysis and on apparent guanylyl cyclase activities were abolished almost completely in the presence of the calmodulin antagonist calmidazolium, whose effect was attenuated by added calmodulin. Thus, a Ca2+/calmodulin-dependent cyclic GMP phosphodiesterase is highly active in synaptic areas of the brain and may prevent elevations of intracellular cyclic GMP levels in activated, NO-producing neurons.     

Redox properties of the Fe3+/Fe2+ couple in Arthromyces ramosus class II peroxidase and its cyanide adduct

The thermodynamics of the one-electron reduction of the ferric heme in free and cyanide-bound Arthromyces ramosus peroxidase (ARP), a class II plant peroxidase, were determined through spectro-electrochemical experiments. The data were compared with those for class III horseradish peroxidase C (HRP) and its cyanide adduct, and were interpreted in terms of ligand binding features, electrostatic effects and solvent accessible surface area of the heme group and of catalytically relevant residues in the heme distal site. The values for free and cyanide-bound ARP (−0.183 and −0.390 V, respectively, at 25 °C and pH 7) are higher than those for HRP and HRP-CN. ARP features an enthalpic stabilization of the ferrous state and a remarkably negative reduction entropy, which are both unprecedented for heme peroxidases. Once the compensatory contributions of solvent reorganization are partitioned from the measured reduction enthalpy, the resulting protein-based value for ARP turns out to be less positive than that for HRP by +10 kJ mol⁻¹. The smaller stabilization of the oxidized heme in ARP most probably results from the less pronounced anionic character of the proximal histidine, and the decreased polarity in the heme distal site as compared with HRP, as indicated by the X-ray structures. The surprisingly negative value for ARP is the result of peculiar reduction-induced solvent reorganization effects.     

Tetrasubstituted cyclopentenone‐based fluorescent chemosensors for the selective detection of Fe3+ and Cu2+ ions

Fluorescent chemosensors based on 4‐hydroxy cyclopentenones were synthesized by the base catalyzed reaction of 1,5‐diphenyl‐pentane‐1,3,5‐trione with benzil and thenil. The molecule obtained by the benzil reaction was found to be useful for the selective detection of Fe³⁺ by fluorescence turn‐off, while the molecule synthesized by the thenil reaction was useful for selective detection of Cu²⁺ by fluorescent turn‐on. Details of the synthesis, complexation mode, nature of binding, reversibility, and pH studies of the two sensors are discussed. The studies revealed that the sensors were suitable for determining Fe³⁺ and Cu²⁺ content in real water samples.     

Tunable warm white light emission and energy transfer of Dy3+ co-doped Sr2LaZrO5.5:Eu3+ phosphors

Eu³⁺,Dy³⁺ co-doped Sr₂LaZrO_5.5-based phosphors were prepared through a sol–gel method. Through characterization, it was found that the Sr₂LaZrO_5.5-based fluorescent powder co-doped with Eu³⁺ and Dy³⁺ had a cubic structure. At an excitation wavelength of 290 nm, the substrate Sr₂LaZrO_5.5 exhibited strong blue emission at 468 nm, and the Sr₂LaZrO_5.5:18%Eu³⁺ phosphor exhibited a strong red emission peak at 612 nm. When the doping amount of Dy³⁺ was 5, 8, 12, 15, or 18%, the Sr₂LaZrO_5.5:18%Eu³⁺ phosphor changed from an orange-red light, to a warm white light, and to a cold white light. According to the emission spectra, the emission intensities of the substrates Sr₂LaZrO_5.5 and Sr₂LaZrO_5.5:Eu³⁺ decreased with increasing Dy³⁺ concentration, confirming the energy transfer between the host Sr₂LaZrO_5.5-Eu³⁺,Dy³⁺, and resulting in a lower CCT value, with significantly improved white light emission.     

Function and Regulation of the Na+-Ca2+ Exchanger NCX3 Splice Variants in Brain and Skeletal Muscle

Isoform 3 of the Na⁺-Ca²⁺ exchanger (NCX3) is crucial for maintaining intracellular calcium ([Ca²⁺]_i) homeostasis in excitable tissues. In this sense NCX3 plays a key role in neuronal excitotoxicity and Ca²⁺ extrusion during skeletal muscle relaxation. Alternative splicing generates two variants (NCX3-AC and NCX3-B). Here, we demonstrated that NCX3 variants display a tissue-specific distribution in mice, with NCX3-B as mostly expressed in brain and NCX-AC as predominant in skeletal muscle. Using Fura-2-based Ca²⁺ imaging, we measured the capacity and regulation of the two variants during Ca²⁺ extrusion and uptake in different conditions. Functional studies revealed that, although both variants are activated by intracellular sodium ([Na⁺]_i), NCX3-AC has a higher [Na⁺]_i sensitivity, as Ca²⁺ influx is observed in the presence of extracellular Na⁺. This effect could be partially mimicked for NCX3-B by mutating several glutamate residues in its cytoplasmic loop. In addition, NCX3-AC displayed a higher capacity of both Ca²⁺ extrusion and uptake compared with NCX3-B, together with an increased sensitivity to intracellular Ca²⁺. Strikingly, substitution of Glu⁵⁸⁰ in NCX3-B with its NCX3-AC equivalent Lys⁵⁸⁰ recapitulated the functional properties of NCX3-AC regarding Ca²⁺ sensitivity, Lys⁵⁸⁰ presumably acting through a structure stabilization of the Ca²⁺ binding site. The higher Ca²⁺ uptake capacity of NCX3-AC compared with NCX3-B is in line with the necessity to restore Ca²⁺ levels in the sarcoplasmic reticulum during prolonged exercise. The latter result, consistent with the high expression in the slow-twitch muscle, suggests that this variant may contribute to the Ca²⁺ handling beyond that of extruding Ca²⁺.     

Secondary Mineral Formation During Ferrihydrite Reduction by Shewanella oneidensis MR-1 Depends on Incubation Vessel Orientation and Resulting Gradients of Cells,Fe2+ and Fe Minerals

In previous studies on microbial ferric iron (Fe(III)) reduction varying results regarding reduction rates and secondary mineral formation have been reported for almost identical conditions regarding temperature, pH, medium composition, Fe(III) mineral identity and bulk iron concentration. Here we show that in addition to physico-chemical parameters also geometric aspects, i.e., incubation orientation and dimension of cultivation vessels, influence the reduction rates and mineralogy. We incubated the Fe(III)-reducer Shewanella oneidensis MR-1 in test tubes at ferrihydrite (FH) concentrations of 1.3–50 mM either in vertical or horizontal orientation. Cells and minerals formed a pellet at the bottom of the tubes with different thicknesses at the same initial FH concentration depending on the incubation orientation. In vertically incubated tubes thick FH pellets were present at the bottom of the tubes and magnetite was formed in all setups with ≥2.5 mM initial FH. In tubes that were incubated horizontally no magnetite was formed in presence of <5 mM initial FH. Spatially resolved analysis of the supernatant and mineral sediment including voltammetric microelectrodes, Xray diffraction and Mössbauer spectroscopy revealed strong gradients of Fe²⁺ in both the aqueous supernatant and mineral pellets, whereas a heterogeneous distribution of cells and minerals in the sediment pellet was detected. The highest cell density and, consequently, the initiation of FH reduction was found at the mineral-supernatant interface. This study demonstrates that small changes in incubation conditions can significantly influence and even change the experimental results of geomicrobiological experiments.     

Regulation by Ca2+ and inositol 1,4,5-trisphosphate (InsP3) of single recombinant type 3 InsP3 receptor channels. Ca2+ activation uniquely distinguishes types 1 and 3 insp3 receptors

The inositol 1,4,5-trisphosphate (InsP(3)) receptor (InsP3R) is an endoplasmic reticulum-localized Ca2+ -release channel that controls complex cytoplasmic Ca(2+) signaling in many cell types. At least three InsP3Rs encoded by different genes have been identified in mammalian cells, with different primary sequences, subcellular locations, variable ratios of expression, and heteromultimer formation. To examine regulation of channel gating of the type 3 isoform, recombinant rat type 3 InsP3R (r-InsP3R-3) was expressed in Xenopus oocytes, and single-channel recordings were obtained by patch-clamp electrophysiology of the outer nuclear membrane. Gating of the r-InsP3R-3 exhibited a biphasic dependence on cytoplasmic free Ca2+ concentration ([Ca2+]i). In the presence of 0.5 mM cytoplasmic free ATP, r-InsP3R-3 gating was inhibited by high [Ca2+]i with features similar to those of the endogenous Xenopus type 1 Ins3R (X-InsP3R-1). Ca2+ inhibition of channel gating had an inhibitory Hill coefficient of approximately 3 and half-maximal inhibiting [Ca2+]i (Kinh) = 39 microM under saturating (10 microM) cytoplasmic InsP3 concentrations ([InsP3]). At [InsP3] < 100 nM, the r-InsP3R-3 became more sensitive to Ca2+ inhibition, with the InsP(3) concentration dependence of Kinh described by a half-maximal [InsP3] of 55 nM and a Hill coefficient of approximately 4. InsP(3) activated the type 3 channel by tuning the efficacy of Ca2+ to inhibit it, by a mechanism similar to that observed for the type 1 isoform. In contrast, the r-InsP3R-3 channel was uniquely distinguished from the X-InsP3R-1 channel by its enhanced Ca2+ sensitivity of activation (half-maximal activating [Ca2+]i of 77 nM instead of 190 nM) and lack of cooperativity between Ca2+ activation sites (activating Hill coefficient of 1 instead of 2). These differences endow the InsP3R-3 with high gain InsP3-induced Ca2+ release and low gain Ca2+ -induced Ca2+ release properties complementary to those of InsP3R-1. Thus, distinct Ca2+ signals may be conferred by complementary Ca2+ activation properties of different InsP3R isoforms.