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Abstract The Aira caldera, located in southern Kyushu, Japan, originally formed 100 ka, and its current shape reflects the more recent 30 ka caldera-forming eruptions (hereafter, called the AT eruptions). This study aimed to delineate the detailed two-dimensional (2D) seismic velocity structure of the Aira caldera down to approximately 15 km, by means of the travel-time tomography analysis of the seismic profile across the caldera acquired in 2017 and 2018. A substantial structural difference in thickness in the subsurface low-velocity areas in the Aira caldera between the eastern and western sides, suggest that the Aira caldera comprises at least two calderas, identified as the AT and Wakamiko calderas. The most interesting feature of the caldera structure is the existence of a substantial high-velocity zone (HVZ) with a velocity of more than 6.8 km/s at depths of about 6–11 km beneath the central area of the AT caldera. Because no high ratio of P- to S-wave velocity zones in the depth range were detected from the previous three-dimensional velocity model beneath the AT caldera region, we infer that the HVZ is not an active magma reservoir but comprises a solidified and cool remnant. In addition, a poorly resolved low-velocity zone around 15 km in depth suggests the existence of a deep active magma reservoir. By superimposing the distribution of the known pressure sources derived from the observed ground inflation and the volcanic earthquake distribution onto the 2D velocity model, the magma transportation path in the crust was imaged. This image suggested that the HVZ plays an important role in magma transportation in the upper crust. Moreover, we estimated that the AT magma reservoir in the 30 ka Aira caldera-forming eruptions has the total volume of 490 km3 DRE and is distributed in a depth range of 4–11 km. Graphical Abstract

Abstract Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) causes coronavirus disease 2019 (COVID-19). Variants of concern (VOCs) such as Delta and Omicron have developed, which continue to spread the pandemic. It has been reported that these VOCs reduce vaccine efficacy and evade many neutralizing monoclonal antibodies (mAbs) that target the receptor binding domain (RBD) of the glycosylated spike (S) protein, which consists of the S1 and S2 subunits. Therefore, identification of optimal target regions is required to obtain neutralizing antibodies that can counter VOCs. Such regions have not been identified to date. We obtained 2 mAbs, NIBIC-71 and 7G7, using peripheral blood mononuclear cells derived from volunteers who recovered from COVID-19. Both mAbs had neutralizing activity against wild-type SARS-CoV-2 and Delta, but not Omicron. NIBIC-71 binds to the RBD, whereas 7G7 recognizes the N-terminal domain of the S1. In particular, 7G7 inhibited S1/S2 cleavage but not the interaction between the S protein and angiotensin-converting enzyme 2; it suppressed viral entry. Thus, the efficacy of a neutralizing mAb targeting inhibition of S1/2 cleavage was demonstrated. These results suggest that neutralizing mAbs targeting blockade of S1/S2 cleavage are likely to be cross-reactive against various VOCs.

Abstract We estimate the mass and energy budgets for the 2018 phreatic eruption of Mt. Motoshirane on Kusatsu–Shirane volcano, Japan, based on data obtained from a network of eight tiltmeters and weather radar echoes. The tilt records can be explained by a subvertical crack model. Small craters that were formed by previous eruptions are aligned WNW–ESE, which is consistent with the strike of the crack modeled in this study. The direction of maximum compressive stress in this region is horizontal and oriented WNW–ESE, allowing fluid to intrude from depth through a crack with this orientation. Based on the crack model, hypocenter distribution, and MT resistivity structure, we infer that fluid from a hydrothermal reservoir at a depth of 2 km below Kusatsu–Shirane volcano has repeatedly ascended through a pre-existing subvertical crack. The inflation and deflation volumes during the 2018 eruption are estimated to have been 5.1 × 105 and 3.6 × 105 m3, respectively, meaning that 1.5 × 105 m3 of expanded volume formed underground. The total heat associated with the expanded volume is estimated to have been ≥ 1014 J, similar to or exceeding the annual heat released from Yugama Crater Lake of Mt. Shirane and that from the largest eruption during the past 130 year. Although the ejecta mass of the 2018 phreatic eruption was small, the eruption at Mt. Motoshirane was not negligible in terms of the energy budget of Kusatsu–Shirane volcano. A water mass of 0.1–2.0 × 107 kg was discharged as a volcanic cloud, based on weather radar echoes, which is smaller than the mass associated with the deflation. We suggest that underground water acted as a buffer against the sudden intrusion of hydrothermal fluids, absorbing some of the fluid that ascended through the crack.

Abstract The tectonic stress field was investigated in and around the aftershock area of the Hokkaido Eastern Iburi earthquake (M JMA = 6.7) occurred on 6 September 2018. We deployed 26 temporary seismic stations in the aftershock area for approximately 2 months and located 1785 aftershocks precisely. Among these aftershocks, 894 focal mechanism solutions were determined using the first-motion polarity of P wave from the temporary observation and the permanent seismic networks of Hokkaido University, Japan Meteorological Agency (JMA), and High Sensitivity Seismograph Network Japan (Hi-net). We found that (1) the reverse faulting and the strike-slip faulting are dominant in the aftershock area, (2) the average trend of P- and T-axes is 78° ± 33° and 352° ± 51°, respectively, and (3) the average plunge of P- and T-axes is 25° ± 16° and 44° ± 20°, respectively: the P-axis is close to be horizontal and the T-axis is more vertical than the average of the P-axes. We applied a stress inversion method to the focal mechanism solutions to estimate a stress field in the aftershock area. As a result, we found that the reverse fault type stress field is dominant in the aftershock area. An axis of the maximum principal stress (σ 1) has the trend of 72° ± 7° and the dipping eastward of 19° ± 4° and an axis of the intermediate principal stress (σ 2) has the trend of 131° ± 73° and the dipping southward of 10° ± 9°, indicating that both of σ 1- and σ 2-axes are close to be horizontal. An axis of the minimum principal stress (σ 3) has the dipping westward of 67° ± 6° that is close to be vertical. The results strongly suggest that the reverse-fault-type stress field is predominant as an average over the aftershock area which is in the western boundary of the Hidaka Collision Zone. The average of the stress ratio R = (σ 1 − σ 2)/(σ 1 − σ 3) is 0.61 ± 0.13 in the whole aftershock area. Although not statistically significant, we suggest that R decreases systematically as the depth is getting deep, which is modeled by a quadratic polynomial of depth.

Abstract Kusatsu-Shirane volcano hosts numerous thermal springs, fumaroles, and the crater lake of Yugama. Hence, it has been a particular study field for hydrothermal systems and phreatic eruptions. On 23 January 2018, a phreatic eruption occurred at the Motoshirane cone of Kusatsu-Shirane, where no considerable volcanic activity had been reported in observational and historical records. To understand the eruption process of this unique event, we analyzed seismic, tilt, and infrasound records. The onset of surface activity accompanied by infrasound signal was preceded by volcanic tremor and inflation of the volcano for ~ 2 min. Tremor signals with a frequency band of 5–20 Hz remarkably coincide with the rapid inflation. We apply an amplitude source location method to seismic signals in the 5–20 Hz band to estimate tremor source locations. Our analysis locates tremor sources at 1 km north of Motoshirane and at a depth of 0.5–1 km from the surface. Inferred source locations correspond to a conductive layer of impermeable cap-rock estimated by magnetotelluric investigations. An upper portion of the seismogenic region suggests hydrothermal activity hosted beneath the cap-rock. Examined seismic signals in the 5–20 Hz band are typically excited by volcano-tectonic events with faulting mechanism. Based on the above characteristics and background, we interpret that excitation of examined volcanic tremor reflects small shear fractures induced by sudden hydrothermal fluid injection to the cap-rock layer. The horizontal distance of 1 km between inferred tremor sources and Motoshirane implies lateral migration of the hydrothermal fluid, although direct evidence is not available. Kusatsu-Shirane has exhibited unrest at the Yugama lake since 2014. However, the inferred tremor source locations do not overlap active seismicity beneath Yugama. Therefore, our result suggests that the 2018 eruption was triggered by hydrothermal fluid injection through a different pathway from that has driven unrest activities at Yugama.

Abstract The Hokkaido Eastern Iburi earthquake (M JMA = 6.7) occurred on September 6, 2018, in the Hokkaido corner region where the Kurile and northeastern Japan island arcs meet. We relocated aftershocks of this intraplate earthquake immediately after the main shock by using data from a permanent local seismic network and found that aftershock depths were concentrated from 20 to 40 km, which is extraordinarily deep compared with other shallow intraplate earthquakes in the inland area of Honshu and Kyushu, Japan. Further, we found that the aftershock area consists of three segments. The first segment is located in the northern part of the aftershock area, the second segment lies in the southern part, and the third segment forms a stepover between the other two segments. The hypocenter of the main shock, from which the rupture initiated, is located on the stepover segment. The centroid moment tensor solution for the main shock indicates a reverse faulting, whereas the focal mechanism solution determined by using the first-motion polarity of the P wave indicates strike-slip faulting. To explain this discrepancy qualitatively, we present a model in which the rupture started as a small strike-slip fault in the stepover segment of the aftershock area, followed by two large reverse faulting ruptures in the northern and southern segments.

Bioluminescence reactions are widely applied in optical in vivo imaging in the life science and medical fields. Such reactions produce light upon the oxidation of a luciferin (substrate) catalyzed by a luciferase (enzyme), and this bioluminescence enables the quantification of tumor cells and gene expression in animal models. Many researchers have developed single-color or multicolor bioluminescence systems based on artificial luciferin analogues and/or luciferase mutants, for application in vivo bioluminescence imaging (BLI). In the current review, we focus on the characteristics of firefly BLI technology and discuss the development of luciferin analogues for high-resolution in vivo BLI. In addition, we discuss the novel luciferin analogues TokeOni and seMpai, which show potential as high-sensitivity in vivo BLI reagents.

This study concerns the multiaxial static and fatigue strength properties. Short-glass-fibre-reinforced phenolic-resin composites (SGP) molded by injection and compression processes were subjected to tensiontorsion combined static and fatigue tests at room temperature under various test conditions. Tension – torsion combined static strength well agreed with Tsai-Hill failure criteria without depending on processes. Relationships between the maximum principal stress, σp1, max, and the number of fracture cycles, Nf, were approximately linear in the whole range of up to 106 cycles. For a unified evaluation of multiaxial fatigue life for SGP, non-dimensional effective stress, σ*, defined by modifying Tsai-Hill failure criteria was applied. The slopes of σ*- Nf curves according to Baskin’s law were almost identical to the injection (n = 26.3) and compression (n = 26.2). We finally confirmed that the multiaxial fatigue life of SFRP could be predicted by using σ* with a unique Wöhler curve without relying on molding processes

Abstract Magnitude–frequency distributions of volcanic explosion earthquakes that are associated with occurrences of vulcanian and strombolian eruptions, or gas burst activity, are examined at six active volcanoes. The magnitude–frequency distribution at Suwanosejima volcano, Japan, shows a power-law distribution

Natural oligonucleotides have many rotatable single bonds, and thus their structures are inherently flexible. Structural flexibility leads to an entropic loss when unwound oligonucleotides form a duplex with single-stranded DNA or RNA. An effective approach to reduce such entropic loss in the duplex-formation is the conformational restriction of the flexible phosphodiester linkage and/or sugar moiety. We here report the synthesis and biophysical properties of a novel artificial nucleic acid bearing an oxanorbornane scaffold (OxNorNA), where the adamant oxanorbornane was expected to rigidify the structures of both the linkage and sugar parts of nucleic acid. OxNorNA phosphoramidite with a uracil (U) nucleobase was successfully synthesized over 15 steps from a known sugar-derived cyclopentene. Thereafter, the given phosphoramidite was incorporated into the designed oligonucleotides. Thermal denaturation experiments revealed that oligonucleotides modified with the conformationally restricted OxNorNA-U properly form a duplex with the complementally DNA or RNA strands, although the Tm values of OxNorNA-U-modified oligonucleotides were lower than those of the corresponding natural oligonucleotides. As we had designed, entropic loss during the duplex-formation was reduced by the OxNorNA modification. Moreover, the OxNorNA-U-modified oligonucleotide was confirmed to have extremely high stability against 3′-exonuclease activity, and its stability was even higher than those of the phosphorothioate-modified counterparts (Sp and Rp). With the overall biophysical properties of OxNorNA-U, we expect that OxNorNA could be used for specialized applications, such as conformational fixation and/or bio-stability enhancement of therapeutic oligonucleotides (e.g., aptamers).

The input of agrochemicals in the aquatic compartment can results in biochemical injuries for living organisms. In this context, the knowledge of alterations of enzymatic activities due the presence of agriculture pollutants contributes for the elucidation of the mechanisms of toxicity, implementation of economic methods for monitoring purposes and establishment of maximum allowed concentrations. In the present work, the above considerations are discussed, and data concerning changes in enzymatic function by pesticides and fertilizer contaminants are reviewed. Also, we focused on the acid phosphatase due its susceptibility to several pollutants and diversity in cellular functions.

Pseudokirchneriella subcapitata is a unicellular green algae widely distributed in freshwater and soils. Due to its cosmopolitan characteristic, its use is recommended by national and international protocols in ecotoxicity studies. The alteration of phosphatase activities by agriculture pollutants like heavy metals has been extensively used as a biomarker in risk assessment and biomonitoring. In this study, we compared the extraction of acid phosphatase from P. subcapitata by different procedures and we studied the stability, substrates specificity, kinetics and the effect of Hg2+ in the crude extract. The freezing and thawing technique associated with probe sonication was the most suitable method of extraction. The enzyme was stable when frozen at -20ºC for at least six months, showed an optimum pH of 5 and a Km value of 0.27 mM for p-nitrophenylphosphate (pNPP) as substrate. Some natural organic substrates were cleaved by a similar extent as the synthetic substrate pNPP. Short term exposure (24 hours) to Hg2+ had little effect but inhibition of the specific activity was observed after 7 days with EC50 (concentration of Hg2+ that promotes 50% decrease of specific activity) value of 12.63 μM Hg2+.Pseudokirchneriella subcapitata é uma alga verde unicelular amplamente distribuída em corpos d´agua e solos. Devido a sua natureza cosmopolita, seu uso é recomendado por protocolos nacionais e internacionais na realização de estudos de ecotoxicidade. A alteração da atividade de fosfatases por agentes poluentes de origem agrícola, como metais pesados, tem sido largamente usada como um biomarcador na avaliação de risco e biomonitoramento. No presente trabalho foi comparada a extração da fosfatase ácida de P. subcapitata por diferentes métodos e estudada a sua estabilidade, especificidade por substratos, cinética e efeito do Hg2+ no extrato bruto. O congelamento e descongelamento, associado com ultrassom, foi o método que proporcionou maior rendimento de extração. A enzima, praticamente estável por armazenamento a -20ºC, durante aproximadamente seis meses, demonstrou uma atividade ótima em pH 5 e um valor de Km para o p-nitrofenilfostato (pNPP) de 0,27 mM. Alguns substratos naturais foram hidrolisados com uma intensidade semelhante à do substrato sintético pNPP. Diferentemente dos estudos de exposição a curto prazo (24 horas), observou-se inibição da atividade específica nas culturas expostas durante 7 dias, com um valor de CE50 (concentração de Hg2+ que promove 50% de decréscimo da atividade específica) equivalente a 12,63 μM Hg2+.

There are few long-term reports of patients with frequent PVCs in the absence of ischemic heart disease. In 86 patients without ischemic heart disease, who had 1000 or more PVCs in 24-hour Holter ECG, the number of PVCs during 24-hours Holter ECG and echocardiographic parameters were followed at least 1 year (66.5 ± 39.7 months). PVC was significantly reduced in the patients with or without underlying diseases (UD). The reduction rate in the number of PVCs was prominent in patients with UD. PVC was significantly reduced in patients under medication, but not in patients without medication. In the comparison between the initial and follow up observation using Wilcoxon's rank test, the number of PVC was significantly reduced (P < 0.05), and EF was also improved (P < 0.05) in angiotensin converting enzyme inhibitor (ACEI) or angiotensin receptor blocker (ARB) group, and in β-blocker group. In Ca-antagonist group and antiarrhythmic drug group, the number of PVCs was also significantly reduced (P < 0.05). Multivariate analysis revealed significantly higher incidence (60% or more with PVC reduction) in ACEI/ARB group. These results suggest that the administration of ACEI/ARB may contribute to the reduction of PVC in non-ischemic heart disease cases with multiple PVC.

Gap junction (GJ) channels provide direct passage for ions and small molecules to be exchanged between neighbouring cells and are crucial for many physiological processes. GJ channels can be gated by transjunctional voltage (known as Vj-gating) and display a wide range of unitary channel conductance (γj), yet the domains responsible for Vj-gating and γj are not fully clear. The first extracellular domain (E1) of several connexins has been shown to line part of their GJ channel pore and play important roles in Vj-gating properties and/or ion permeation selectivity. To test roles of the E1 of Cx50 GJ channels, we generated a chimera, Cx50Cx36E1, where the E1 domain of Cx50 was replaced with that of Cx36, a connexin showing quite distinct Vj-gating and γj from those of Cx50. Detailed characterizations of the chimera and three point mutants in E1 revealed that, although the E1 domain is important in determining γj, the E1 domain of Cx36 is able to effectively function within the context of the Cx50 channel with minor changes in Vj-gating properties, indicating that sequence differences between the E1 domains in Cx36 and Cx50 cannot account for their drastic differences in Vj-gating and γj. Our homology models of the chimera and the E1 mutants revealed that electrostatic properties of the pore-lining residues and their contribution to the electric field in the pore are important factors for the rate of ion permeation of Cx50 and possibly other GJ channels.

Deprotonation or suppression of the pKa of the amino group of a lysine sidechain is a widely recognized phenomenon whereby the sidechain amino group transiently can act as a nucleophile at the active site of enzymatic reactions. However, a deprotonated lysine and its molecular interactions have not been directly experimentally detected. Here, we demonstrate a deprotonated lysine stably serving as an "acceptor" in a H-bond between the photosensor protein RcaE and its chromophore. Signal splitting and trans-H-bond J coupling observed by NMR spectroscopy provide direct evidence that Lys261 is deprotonated and serves as a H-bond acceptor for the chromophore NH group. Quantum mechanical/molecular mechanical calculations also indicate that this H-bond exists stably. Interestingly, the sidechain amino group of the lysine can act as both donor and acceptor. The remarkable shift in the H-bond characteristics arises from a decrease in solvation, triggered by photoisomerization. Our results provide insights into the dual role of this lysine. This mechanism has broad implications for other biological reactions in which lysine plays a role. [ABSTRACT FROM AUTHOR]

We tried infrasonic monitoring of snow avalanche at Mt. Fuji. We installed infrasound sensors, a thermometer-probe snow-depth meter, and seismometers for two winter seasons from 2018 to 2020 and made integrated analyses of their data. Although no significant snow avalanche event occurred during our observation, we found many infrasound waveforms similar to the reported snow avalanche infrasound. Because the infrasound events accompanied little seismic power, we inferred surface phenomena, if not necessarily snow avalanches, generated them. The events' occurrences were concentrated during and right after snow-falls and during snow melting. The infrasonic array analyses of the 2019-2020 dataset exhibited most of the events are from the north slopes below the observation site. From the summit direction, continuous infrasonic noise was frequently detected, by which detecting events from there might have been difficult. This study has shown that the multi-parameter observation method is practical and useful in the severe environment of Mt. Fuji in winter. Also, it revealed the particular problems of infrasound observation at the huge volcanic edifice. We expect our results will help design future observations. [ABSTRACT FROM AUTHOR]

Cytochrome c oxidase (CcO), a membrane enzyme in the respiratory chain, catalyzes oxygen reduction by coupling electron and proton transfer through the enzyme with a proton pump across the membrane. In all crystals reported to date, bovine CcO exists as a dimer with the same intermonomer contacts, whereas CcOs and related enzymes from prokaryotes exist as monomers. Recent structural analyses of the mitochondrial respiratory supercomplex revealed that CcO monomer associates with complex I and complex III, indicating that the monomeric state is functionally important. In this study, we prepared monomeric and dimeric bovine CcO, stabilized using amphipol, and showed that the monomer had high activity. In addition, using a newly synthesized detergent, we determined the oxidized and reduced structures of monomer with resolutions of 1.85 and 1.95 Å, respectively. Structural comparison of the monomer and dimer revealed that a hydrogen bond network of water molecules is formed at the entry surface of the proton transfer pathway, termed the K-pathway, in monomeric CcO, whereas this network is altered in dimeric CcO. Based on these results, we propose that the monomer is the activated form, whereas the dimer can be regarded as a physiological standby form in the mitochondrial membrane. We also determined phospholipid structures based on electron density together with the anomalous scattering effect of phosphorus atoms. Two cardiolipins are found at the interface region of the supercomplex. We discuss formation of the monomeric CcO, dimeric CcO, and supercomplex, as well as their role in regulation of CcO activity. [ABSTRACT FROM AUTHOR]

The detection capability of various anomalous phenomena preceding volcanic eruptions has considerably progressed as the geophysical monitoring networks have become denser and multi-disciplinary. However, current eruption forecasting techniques, from a practical perspective, still have much scope for improvement because they largely depend on empirical techniques. In the past decade, threedimensional modeling based on the electromagnetic sounding methods such asmagnetotellurics (MT) have become a practical choice, and its recent applications to active volcanic fields has revealed certain common features among volcanoes. Information about the resistivity structure, especially in 'wet' volcanic fields, is useful for the provisional screening of the eruption potential from the viewpoint of the subsurface structure, and, thus, may contribute to the evaluation of eruption imminence in a broad sense. In this study, for evaluation purposes, we present the roles and possible further applications of the subsurface resistivity structure studies by demonstrating the preliminary results and interpretations of an MT survey that we performed in the Kuttara Volcanic Group, northern Japan. [ABSTRACT FROM AUTHOR]

A new approach to platencin, a potent antibiotic isolated from Streptomyces platensis, has been established. The highly congested tricyclic core of the natural product was successfully constructed by decarboxylative radical cyclization of an alkynyl silyl ester with Pb(OAc)4 in the presence of pyridine in refluxing 1,4-dioxane. The key decarboxylation, which likely takes place via lead(IV) esterification followed by carbon-centered radical generation and subsequent capture of the radical with a triple bond, allows the rapid construction of the twisted polycyclic system. [ABSTRACT FROM AUTHOR]

A 47‐year‐old man with drug‐resistant paroxysmal atrial fibrillation underwent left atrial (LA) catheter ablation. After sheaths were inserted into the LA using the Brockenbrough method, he complained of angina and developed ST‐segment elevation in the II, III, aVF, V5, and V6 leads. Ventricular fibrillation (VF) occurred followed by ventricular tachycardia. The VF was successfully treated with direct current shock. Coronary angiography with isosorbide dinitrate showed neither spasm nor embolism in the coronary artery. The chest pain disappeared after 6 min with resolution of the ST‐segment elevation. A summarized review of ST‐segment elevation associated with transseptal puncture disclosed that ST‐segment elevation is often found in inferior leads (87.5%), while concomitant bradyarrhythmia has been reported in 43.8% of patients. Our patient is the first recorded case with potentially lethal tachyarrhythmia. Although ST‐segment elevation and VF is a rare complication associated with transseptal puncture, awareness of this complication is important. [ABSTRACT FROM AUTHOR]

Abstract  The presence of copper in water environment may have detrimental effects on aquatic organisms, including algae, where different enzymatic systems can be affected. Algae acid phosphatase plays important roles in metabolic processes such as decomposition of organic phosphate, autophagic digestive process, recycling cellular materials and zygote formation during reproduction. This work describes an in vitro activation effect of copper on the acid phosphatase of the green algae Pseudokirchneriella subcapitata (formely Selenastrum capricornutum) under preincubation condition. Apparent Michaelis constant values of 1.21 and 0.37 mM, and activation energy values of 26.8 and 13.6 kJ mol−1 were determined in the absence and in the presence of 0.2 mM Cu2, respectively. The dissociation constant value for Cu2 binding to the enzyme was determined to be 22.04 μM. The decrease of the apparent Michaelis constant (Km) and activation energy values in the presence of Cu2 correlates well with its activating effect on the acid phosphatase activity. This propriety could be used as a sensitive bioindicator for copper in environmental samples. [ABSTRACT FROM AUTHOR]

The effect of temperature on the activity and structural stability of an acid phosphatase (EC 3.1.3.2.) purified from castor bean (Ricinus communis L.) seeds have been examined. The enzyme showed high activity at 45 °C using p-nitrophenylphosphate (p-NPP) as substrate. The activation energy for the catalyzed reaction was 55.2 kJ mol−1 and the enzyme maintained 50% of its activity even after 30 min at 55 °C. Thermal inactivation studies showed an influence of pH in the loss of enzymatic activity at 60 °C. A noticeable protective effect from thermal inactivation was observed when the enzyme was preincubated, at 60 °C, with the reaction products inorganic phosphate—P (10 mM) and p-nitrophenol—p-NP(10 mM). Denaturation studies showed a relatively high transition temperature (Tm) value of 75 °C and an influence of the combination of Pi (10 mM) and p-NP (10 mM) was observed on the conformational behaviour of the macromolecule (Mol Cell Biochem 266: 11–15, 2004) [ABSTRACT FROM AUTHOR]

The effect of anionic and cationic surfactants on acid phosphatase denaturation has been extensively studied. Low molecular mass (LMr) protein tyrosine phosphatase (PTP), a key regulatory enzyme involved in many different processes in the cell, was distinctly affected by anionic (homologous series of n-alkyl sulfates (C8-C14)) and cationic (n-alkyl trimethylammonium bromides (C12-C16)) surfactants. At concentrations 10-fold lower critical micellar concentration (cmc) values, the enzyme was completely inactivated in the presence of anionic surfactants, in a process independent of the pH, and dependent on the chain length of the surfactants. Under the same conditions, the effect of cationic surfactants on the enzyme activity was pH-dependent and only at pH 7.0 full inactivation was observed at concentrations 10-fold higher cmc values. In contrast to cationic surfactants the effect of anionic surfactants on the enzyme activity was irreversible and was not affected by the presence of NaCl. Inorganic phosphate, a known competitive inhibitor of PTP, protected the enzyme against inactivation by the surfactants. Our results suggest that the inactivation of the LMr PTP by anionic and cationic surfactants involved both electrostatic and hydrophobic interactions, and that the interactions enzyme-surfactants probably occurred at or near the active site. (Mol Cell Biochem 265: 133–140, 2004) [ABSTRACT FROM AUTHOR]

An acid phosphatase (EC 3.1.3.2) has been identified and purified from castor bean (Ricinus communis L., IAC-80) seed through sulphopropyl (SP)-Sephadex, diethylaminoethyl (DEAE)-Sephadex, Sephacryl S-200, and Concanavalin A-Sepharose chromatography. The enzyme was purified 2 000-fold to homogeneity, with a final specific activity of 3.8 μkat mg-1 protein. The purified enzyme revealed a single diffuse band with phosphatase activity on nondenaturing polyacrylamide gel electrophoresis, at pH 8.3. The relative molecular mass, determined by high-performance liquid chromatography (HPLC), was found to be 60 kDa. The acid phosphatase had a pH optimum of 5.5 and an apparent Km value for p-nitrophenylphosphate of 0.52 mM. The enzyme-catalyzed reaction was inhibited by inorganic phosphate, fluoride, vanadate, molybdate, p-chloromercuribenzoate (pCMB), Cu2+ and Zn2+. The strong inhibition by pCMB, Cu2+ and vanadate suggests the presence of sulfhydryl groups essential for catalysis. The castor bean enzyme also recognized tyrosine-phosphate and inorganic pyrophosphate (PPi) as substrate. The highest specificity constant (Vmax/Km) was observed with PPi, making it a potential physiological substrate. [ABSTRACT FROM AUTHOR]

Certain cyanobacteria alter their photosynthetic light absorption between green and red, a phenomenon called complementary chromatic acclimation. The acclimation is regulated by a cyanobacteriochrome-class photosensor that reversibly photoconverts between green-absorbing (Pg) and red-absorbing (Pr) states. Here, we elucidated the structural basis of the green/red photocycle. In the Pg state, the bilin chromophore adopted the extended C15-Z,anti structure within a hydrophobic pocket. Upon photoconversion to the Pr state, the bilin is isomerized to the cyclic C15-E,syn structure, forming a water channel in the pocket. The solvation/desolvation of the bilin causes changes in the protonation state and the stability of π-conjugation at the B ring, leading to a large absorption shift. These results advance our understanding of the enormous spectral diversity of the phytochrome superfamily. [ABSTRACT FROM AUTHOR]