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1.
HP1454 is a protein of 303 amino acids found in the extracellular milieu of Helicobacter pylori. The protein structure, crystallized in the orthorhombic C2221 space group with one molecule per asymmetric unit, has been determined using the single‐wavelength anomalous dispersion method. HP1454 exhibits an elongated bent shape, composed of three distinct domains. Each domain possesses a fold already present in other structures: Domain I contains a three‐strand antiparallel β‐barrel flanked by a long α‐helix, Domain II is an anti‐parallel three‐helix bundle, and Domain III a β‐sheet flanked by two α‐helices. The overall assembly of the protein does not bear any similarity with known structures. Proteins 2014; 82:2868–2873. © 2014 Wiley Periodicals, Inc.  相似文献   

2.
Diffusion coefficients for thirteen lipophilic organic compounds in reconstituted waxes of Fagus sylvatica L. and Picea abies (L.) Karst. were measured to characterise the transport properties of the cuticular waxes that form the transport-limiting barrier of plant cuticles. Desorption kinetics (relative amounts desorbed versus time) were asymptotic, but could be linearized up to 50% desorption by plotting relative amounts desorbed versus the square root of time. Diffusion coefficients calculated from the slopes of the linear regressions ranged from 10–22 to 10–17 m2·s–1 and decreased with increasing molecular size. This size dependency of diffusion coefficients was analysed, assuming an exponential dependence of the diffusion coefficients on molar volumes, which allowed cuticular transport properties to be related to the physical structure of the wax. Furthermore, the fact that the barrier properties of Fagus wax are less pronounced than those of Picea is interpreted as an ecological adaptation of the respective tree species to their habitats at the level of their cuticular transport barriers.Abbreviation D diffusion coefficient The authors gratefully acknowledge financial support by the Schwerpunkt Programm Baumphysiologie of the Deutsche Forschungsgemeinschaft.  相似文献   

3.
The antifungal plant defensin RsAFP2 isolated from radish interacts with fungal glucosylceramides and induces apoptosis in Candida albicans. To further unravel the mechanism of RsAFP2 antifungal action and tolerance mechanisms, we screened a library of 2868 heterozygous C. albicans deletion mutants and identified 30 RsAFP2‐hypersensitive mutants. The most prominent group of RsAFP2 tolerance genes was involved in cell wall integrity and hyphal growth/septin ring formation. Consistent with these genetic data, we demonstrated that RsAFP2 interacts with the cell wall of C. albicans, which also contains glucosylceramides, and activates the cell wall integrity pathway. Moreover, we found that RsAFP2 induces mislocalization of septins and blocks the yeast‐to‐hypha transition in C. albicans. Increased ceramide levels have previously been shown to result in apoptosis and septin mislocalization. Therefore, ceramide levels in C. albicans membranes were analysed following RsAFP2 treatment and, as expected, increased accumulation of phytoC24‐ceramides in membranes of RsAFP2‐treated C. albicans cells was detected. This is the first report on the interaction of a plant defensin with glucosylceramides in the fungal cell wall, causing cell wall stress, and on the effects of a defensin on septin localization and ceramide accumulation.  相似文献   

4.
Ma  C.C.  Gao  Y.B.  Guo  H.Y.  Wang  J.L. 《Photosynthetica》2004,42(1):65-70
In the order C. microphylla — C. intermedia — C. korshinskii, compensation irradiance, saturation irradiance, and optimum temperature for photosynthesis increased, net photosynthetic rate (P N) at low irradiance and low temperature decreased, optimum air humidity decreased, and P N at low air humidity increased. Daily cumulative value of P N increased while daily cumulative value of transpiration (E) decreased, and hence water use efficiency (WUE =P N/E) increased. Diurnal course of P N of C. microphylla was a double-peak curve, but the second peak in the curves of C. intermedia and C. korshinskii was not visible. These physiological characteristics are biological basis for the geographical distribution of these three Caragana species, and are in relation to water conditions of their habitats and distinctiveness in leaf hair of plant.  相似文献   

5.
The stereochemistry of sulfate conjugation of isoproterenol (ISO) was examined with human liver, intestine, and platelets as the phenolsulfotransferase (PST) enzyme source and PAP35S as the cosubstrate. With the hepatic cytosol, two distinct sulfation reactions were identified, a high affinity reaction (Km 5 to 50 μM) and a low affinity reaction (Km 360 to 2,900 μM). The efficiency of sulfation (Vmax/Km) for both reactions was 5-fold higher for (+)- than for (?)-ISO. When the hepatic PSTs were resolved by ionexchange chromatography, it could be shown that the high affinity reaction was catalyzed by the monoamine (M) form and the low affinity reaction by the phenol (P) form of PST. Only the high affinity (M form) sulfation was detected in the jejunal cytosol with a Vmax/Km value 6.1-fold higher for (+)- than for (?)-ISO. Finally the platelet, as a potentially useful model tissue, also demonstrated only the high affinity M form reaction with a Vmax/Km value 5.7-fold higher for (+)- than for (?)-ISO. In summary, this study has shown that sulfation of ISO by PSTs in various human tissues is stereoselective and favors the inactive (+)-enantiomer over the active (?)-enantiomer by about 5-fold, a finding which should be considered in the therapeutic use of chiral drugs cleared by sulfate conjugation. © 1993 Wiley-Liss, Inc.  相似文献   

6.
Wang  R.Z.  Yuan  Y.Q. 《Photosynthetica》2001,39(2):283-287
The intra- and inter-specific variations in net photosynthetic (P N) and transpiration (E) rates and water use efficiency (WUE) of Puccinellia tenuiflora and Puccinellia chinampoensis leaves were compared. The two species experienced a similar habitat, but differed in leaf area, leaf colour, and nitrogen contents. Leaf P N and E for both reproductive and vegetative shoots of the two species declined with leaf age. P N for reproductive shoots was less than for vegetative shoots, but their E was greater than that of vegetative shoots in the dry season. The average P N and E for reproductive shoots of P. tenuiflora were lower than those of P. chinampoensis, but higher for vegetative shoots.  相似文献   

7.
The subcellular distributions of potassium, sodium, magnesium, calcium and chloride have been determined for rabbit cerebral cortex. After homogenization and differential centrifugation, the following percentages of ions were associated with the particulate fraction (nuclear, mitochondrial, synaptic vesicles, and microsomal): (a) 19% of the total potassium; (b) 22% of the total sodium; (c) 77% of the total calcium; (d) 69% of the total magnesium; and (e) less than 2% of the total chloride. However, the sum of the potassium and sodium content in each of the particulate fractions was greater than the sum of the calcium and magnesium content. After hypo-osmotic shock of the crude mitochondrial fraction (MT), more sodium than potassium (μmol/g wet wt.) was associated with the mitochondrial (M1) and synaptic vesicle (M2) fractions. The molar ratio of sodium to potassium was 1·4 for M1 and 4·5 for M2. The association of 22Na+ with the particulate fractions was further studied by the method of equilibrium dialysis. The data from both types of experiments indicate that a large fraction of the sodium in cortical tissue appears to be in a bound state.  相似文献   

8.
Summary The NADP-specific malate dehydrogenase isozymes were controlled by multiple gene systems. Three genes coding for dimeric enzymes segregated in a dependent fashion (NADP-Mdh 1, NADP-Mdh 2, NADP-Mdh 3). A fourth gene (NADP-Mdh 4), also coded for dimers, but was not polymorphic in B. vulgaris. A fifth gene (NADP-Me 1) coded for enzymes active as monomers. Two genes were found to control the main zone of NAD-specific malate dehydrogenase: one coded for dimers (Mdh 1), while a second (Mdh 2) was not polymorphic in the assessions studied. 6-P-Gluconate dehydrogenase was not polymorphic in B. vulgaris; the two types detected on SGE1 electrophoresis were due to developmental expression of the different systems. No genetical segregations could be detected in progeny of crosses of the distinct phenotypes. A shikimate dehydrogenase gene (Skdh 1) that coded for monomers was identified. The diaphorase system was rather complex, but one gene (Dia 1) coding for monomeric enzymes could be identified. Aconitase was found to be controlled by two independent genes (Aco 1, Aco 2), both polymorphic and coding for proteins active as monomers. Tight linkage was found between the genes NADP-Mdh 1, NADP-Mdh 2 and NADP-Mdh 3. Linkage was also found between a pollen fertility restorer (Z) and the Mdh 1 gene. The identification of linkage with Aco 1 needs further investigation. R segregated independently from Mdh 1, Aco 1 and Dia 1. Independent segregations were scored for isozyme genes Pgm 2, Icd 1, Ak 1, Gpi 1, Aco 1 and Dia 1.Abbreviations Tris-HCl Tris (hydroxymethyl) aminomethane-HCl - NADP nicotinamide adenine dinucleotide phosphate - NBT nitro-blue tetrazolium chloride monohydrate - PMS phenazine methosulphate  相似文献   

9.
Soybean cyst nematode (SCN; Heterodera glycines Ichinohe) is one of the most destructive pests in the cultivation of soybean (Glycine max (L.) Merr.) worldwide. Markers based on the SCN resistance gene will enable efficient marker-assisted selection (MAS). We sequenced the candidate gene rhg1 in six resistant and two susceptible soybean genotypes and identified 37 SNPs (single nucleotide polymorphisms) among the sequences, of which 11 were in the coding region. Seven of these 11 SNPs led to changes in the amino acid sequence of the gene. The amino acid sequence we obtained differs from the previously published one by a stretch of 26–27 amino acids. Six codominant allele-specific SNP markers based on agarose gel detection were developed and tested in 70 genotypes, among which occurred only nine different haplotypes. Two neutrality tests (Tajima’s D and Fu and Li’s F) were significant for the six SNP loci in the 70 genotypes, which is consistent with intensive directional selection. A strong LD pattern was detected among five SNPs except 2868T > C. Two SNPs (689C > A and 757C > T) formed one haplotype (689C-757C) that was perfectly associated with SCN resistance. The new allele-specific PCR markers located in the alleged sequence of the rhg1 candidate gene, combined with the microsatellite marker BACR-Satt309, will significantly improve the efficiency of MAS during the development of SCN-resistant cultivars.  相似文献   

10.
Relationships between number of eggs (Ng), brood weight (Wg), and body length (L) and weight (W) of females were studied for 16 orders and suborders of Crustacea. The study was based on the authors' observations in the inland waters of the European USSR, in the coastal regions of the fareastern and northern seas and data from V. V. KUZNETSOV (preserved in archives). An extensive literature was also used. The equations describing the relationships (Ng=f(L) and Wg=f(W) are calculated for some species and high taxons. On the average for the superclass (excluding the order Notostraca) the Wg/W ratio appeared to be 0.16. This ratio is markedly different for species of Notostraca. It is supposed that this difference is due to the high metabolic rate of Notostraca.  相似文献   

11.
An α-amylase secreted by Pichia burtonii 15-1 isolated from a traditional starter murcha of Nepal, named Pichia burtonii α-amylase (PBA), was studied. The gene was cloned and its nucleotide sequence was determined. PBA was deduced to consist of 494 amino acid residues. It shared certain degrees of amino acid sequence identity with other homologous proteins: 60% with Schwanniomyces occidentalis α-amylase, 58% with Saccharomycopsis sp. α-amylase, and 47% with Taka-amylase A from Aspergillus oryzae. A three-dimensional structural model of PBA generated using the known three-dimensional structure of Taka-amylase A as a template suggested high structural similarity between them. Kinetic analysis revealed that the Km values of PBA were lower than those of Taka-amylase A for the oligosaccharides. Although the kcat values of PBA were lower than those of Taka-amylase A for the oligosaccharide substrates, the kcat/Km values of PBA were higher.  相似文献   

12.
Catalase (CATpp) with molecular weight 223 kD was isolated from the methylotrophic yeast Pichia pastoris and purified 90-fold by ion-exchange chromatography and gel filtration. Quantitative parameters of absorption and CD spectra of CATpp solutions and of its membrane-concentrated form (CATpp-conc) were studied. Rates of H2O2 decomposition and kinetic characteristics K m and k cat of CATpp and CATpp-conc were determined in 10 mM phosphate buffer (pH 7.4) at 30°C, as well as the effective constant k in of the enzyme inactivation rate during the catalysis and the constant k 2 of the interaction rate of the Complex I catalases with H2O2. Thermal inactivation of CATpp in solutions at 45°C was characterized by the effective rate constant k in *, and the low-frequency (27 kHz) ultrasonic inactivation of CATpp at 20°C was characterized by the firstorder rate constant k in (US). All spectral and kinetic characteristics of CATpp and CATpp-conc were compared with the corresponding values for catalase from bovine liver (CAT) and for catalase from the methylotrophic yeast Candida boidinii (CATcb). All three catalases were rather similar in their spectral properties but strongly varied in their kinetic parameters, and their comparison suggests that CATpp should be the best enzyme in its overall properties as it displayed the maximal efficiency in terms of k cat/K m, thermal stability comparable with the thermal stability of CAT in terms of k in *, the minimal k in, and high stability in the ultrasonic cavitation field at the US power of 60 W/cm2.  相似文献   

13.
Age, growth, and age at sexual maturity of the polkadot skate Dipturus chinensis, in the northern East China Sea were determined for a total of 614 specimens collected from April 2009 to December 2014. Vertebral centrum analysis was used to calculate the age of the skates. Annual band deposition was determined by marginal increment analysis. The von Bertalanffy growth model was fitted to the observed length‐at‐age data for each sex (males, L = 76.8, k = 0.109, t0 = ?1.28; females, L = 83.1, k = 0.103, t0 = ?1.20). Growth patterns of females and males were similar until the age of 6; thereafter, females grew larger than males. Maximum age recorded was 13 years for males and 15 years for females. Age at 50% sexual maturity was 8.22 years for males and 9.39 years for females. These results indicate that Dchinensis is slow growing, relatively long‐lived, and late maturing, and therefore vulnerable to exploitation.  相似文献   

14.
The effect of temperature, light-spectrum, desiccation and salinity gradients on the photosynthesis of a Japanese subtidal brown alga, Sargassum macrocarpum (Fucales), was determined using a pulse amplitude modulation-chlorophyll fluorometer and dissolved oxygen sensors. Temperature responses of the maximum (Fv/Fm in darkness) and effective (ΔF/Fm at 50 μmol photons m−2 s−1; = ΦPSII) quantum yields during 6-day culture (4–36°C) remained high at 12–28°C, but decreased at higher temperatures. Nevertheless, ΔF/Fm also dropped at temperatures below 8°C, suggesting light sensitivity under chilling temperatures because Fv/Fm remained high. Photosynthesis–irradiance responses at 24°C under red (660 nm), green (525 nm), blue (450 nm) and white light (metal halide lamp) showed that maximum net photosynthesis under blue and white light was greater than under red and green light, indicating the sensitivity and photosynthetic availability of blue light in the subtidal light environment. In the desiccation experiment, samples under aerial exposure of up to 8 h under dim-light at 24°C and 50% humidity showed that ΔF/Fm quickly declined after more than 45 min of emersion; furthermore, ΔF/Fm also failed to recover to initial levels even after 1 day of rehydration in seawater. Under the emersion state, the ΔF/Fm remained high when the relative water content (RWC) was greater than 50%; in contrast, it quickly dropped when the RWC was less than 50%. When the RWC was reduced below 50%, ΔF/Fm did not return to initial levels, regardless of subsequent re-hydration, suggesting a low capacity of photosynthesis to recover from desiccation. The stenohaline response of photosynthesis under 3-day culture is evident, given that ΔF/Fm declined when salinity was beyond 20–40 psu. Adaptation to subtidal environments in temperate waters of Japan can be linked to these traits.  相似文献   

15.
Studied were the length‐weight relationships (LWRs) and condition factors of two threatened species, Rasbora tawarensis and Poropuntius tawarensis, endemic to Lake Laut Tawar. The objective of the present study was to establish baseline data on LWRs of the two endemic fishes. The growth pattern for both species was predicted using a Linear Allometric Model (LAM) of length‐weight relationship and Fulton’s condition factor (KTL) as well as Relative Weight (Wr). The b value of R. tawarensis was relatively similar between females and males, while the b value of female P. tawarensis was slightly higher than in males. Nevertheless, both R. tawarensis and P. tawarensis showed an allometric negative growth pattern. The average KTL value of R. tawarensis was 1.85 and 1.73 for females and males, respectively, while P. tawarensis had values of 2.44 and 2.41 for females and males, respectively. In addition, the average Wr values of R. tawarensis were 98.9 and 100.45 for females and males respectively, while 102.30 for female and 102.96 for male P. tawarensis, indicating that the condition factors of P. tawarensis were higher than in R. tawarensis.  相似文献   

16.
Xu  Hui-Lian  Gauthier  L.  Desjardins  Y.  Gosselin  A. 《Photosynthetica》1997,33(1):113-123
Gross photosynthetic capacity (P G ) of greenhouse-grown tomato plants (Lycopersicon esculentum Mill.) decreased as the leaf aged. The P G of the 10th, 15th and 18th leaves from the top was only 76, 37, and 18 % of P G of the 5th leaf, respectively. Quantum yield (Y Q ) and dark respiration rate (R D ) were also lower in older leaves than in the younger ones. Net photosynthetic rate (P G ) was apparent in young fruits (about 10 g FM) or young petioles but no P N was found in large fruits (40 g or more FM) and stems because of high R D . Both P G and R D were lower in older fruits and petioles or in lower parts of the stem compared to the younger ones or upper parts of stem. A sharp decrease in chlorophyll (Chl) content was only measured in the senescing 18th leaf. The Chl content in petioles, stems and fruits was proportional to P G . Decreases in P G of older leaves were attributed to decreases in content rather than activity of ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBPCO) since soluble protein content was lower in older leaves than in the younger ones but the specific activity (activity per unit of protein) of RuBPCO was not so. The estimated values of P N of the 10th, 15th and 18th leaves inside the canopy were only 50, 21, and 7 % of that in the 5th leaf. Therefore, leaves below the 18th can be removed in order to ensure a good air circulation and prevent diseases. The significance of photosynthesis in fruit, stem and petioles is not negligible because photosynthesis re-fixes the respired CO2. This revised version was published online in September 2006 with corrections to the Cover Date.  相似文献   

17.
《MABS-AUSTIN》2013,5(8):1322-1331
ABSTRACT

Manipulation of binding affinity between monoclonal antibodies (mAbs) and the neonatal Fc receptor (FcRn) has been leveraged to extend mAb half-life; however, the steps required for success remain ambiguous and experimental observations are inconsistent. Recent models have considered the time course of endosomal transit a major contributor to the relationship between FcRn affinity and antibody half-life. Our objective was to develop a minimal physiologically based pharmacokinetic model to explain how changes in IgG-FcRn association rate constant (Kon), dissociation rate constant (Koff), and endosomal transit time [T(w)] translate to improved IgG clearance across mice, monkeys and humans. By simulating mAb clearance across physiological values of Kon, Koff, and T(w), we found that lowering Koff improves clearance only until the dissociation half-life reaches endosomal transit time. In contrast, Kon influenced clearance independently of T(w).The model was then applied to fit 66 mAb plasma profiles across species digitized from the literature, and clearance of mAb (CLIgG) and vascular fluid-phase endocytosis rate (CLup) were estimated. We found that CLIgG scaled well with body weight (allometric exponent of 0.90). After accounting for mAbs with significant FcRn binding at physiological pH, CLup was allometrically scalable (exponent 0.72). For the antibodies surveyed, Kon was more highly correlated with CLIgG across all species. The relationship between Koff and KD with CLIgG was largely inconsistent. Taken together, this model provides a parsimonious approach to evaluate endosomal transit kinetics using only mAb plasma concentrations. These findings reinforce the idea that endosomal transit kinetics should be considered when modeling FcRn salvage.  相似文献   

18.
This paper describes the modern enzymology in Japanese bioindustries. The invention of Takadiastase by Jokiti Takamine in 1894 has revolutionized the world of industrial enzyme production by fermentation. In 1949, a new γ-amylase (glucan 1,4-α-glucosidase, EC 3.2.1.3) from A. luchuensis (formerly designated as A. awamori), was found by Kitahara. RNase T1 (guanyloribonuclease, EC 3.1.27.3) was discovered by Sato and Egami. Ando discovered Aspergillus nuclease S1 (single-stranded nucleate endonuclease, EC 3.1.30.1). Aspergillopepsin I (EC 3.4.23.18) from A. tubingensis (formerly designated as A. saitoi) activates trypsinogen to trypsin. Shintani et al. demonstrated Asp76 of aspergillopepsin I as the binding site for the basic substrate, trypsinogen. The new oligosaccharide moieties Man10GlcNAc2 and Man11GlcNAc2 were identified with α-1,2-mannosidase (EC 3.2.1.113) from A. tubingensis. A yeast mutant compatible of producing Man5GlcNAc2 human compatible sugar chains on glycoproteins was constructed. The acid activation of protyrosinase from A. oryzae at pH 3.0 was resolved. The hyper-protein production system of glucoamylase was established in a submerged culture.  相似文献   

19.
The complex piezoelectric constant (d = d′ ? id″), elastic constant (c = c′ + ic″), and dielectric constant (ε = ε′ ? iε″) were measured at a frequency of 10 Hz over the temperature range from ?150 to 50°C and for a range of hydration up to 0.26 g/g for decalcified bone and up to 0.084 g/g for bone. For decalcified bone, ε′ and ε″ increased with increasing hydration with a deflection at the critical hydration hc = 0.08 g/g;d′ at ?150°C increased below hc but decreased above hc with increasing hydration; c′ increased below ?60°C but decreased above ?60°C with increasing hydration; and the peak temperatures of ε″, d″, and c″ below ?50°C agree with each other and decreased with increasing hydration with a deflection at hc. For bone, similar hydration and temperature dependences were observed for ε and c. However, the dependence of d on hydration and temperature was different from that of decalcified bone, reflecting a two-phase structure consisting of collagen fibers and mineral hydroxyapatite. The critical hydration for bone was 0.04 g/g.  相似文献   

20.
Lipoprotein-associated phospholipase A2 (Lp-PLA2) has been shown to play a crucial role in atherosclerosis, and has been proposed as a promising target for drug discovery. Here, we cloned the Lp-PLA 2 gene from differentiated THP-1 cells, and inserted a carboxy-terminal His6-tagged version of the gene into the pPIC9 Pichia expression vector. The Lp-PLA2 fusion protein was successfully expressed in Pichia pastoris expression system and could be rapidly purified to apparent homogeneity using a single-step purification method. The activity of our recombinant Lp-PLA2 was strong when [3H] PAF was used as a substrate, and the Lp-PLA2 inhibitor SB435495 exhibited an inhibitory curve against the recombinant Lp-PLA2 (IC50=15.93±1 μM). This novel recombinant Lp-PLA2 could prove useful as a screening model for Lp-PLA2 inhibitors, and may facilitate further investigation of this protein in atherosclerosis.  相似文献   

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