Identification of a Mn-O-Mn cluster vibrational mode of the oxygen-evolving complex in photosystem II by low-frequency FTIR spectroscopy

We have developed conditions for recording the low-frequency S(2)/S(1) Fourier transform infrared difference spectrum of hydrated PSII samples. By exchanging PSII samples with buffered (18)O water, we found that a positive band at 606 cm(-)(1) in the S(2)/S(1) spectrum in (16)O water is clearly downshifted to 596 cm(-)(1) in (18)O water. By taking double-difference (S(2)/S(1) and (16)O minus (18)O) spectra, we assign the 606 cm(-)(1) mode to an S(2) mode and also identify a corresponding S(1) mode at about 625 cm(-)(1). In addition, by Sr and (44)Ca substitution experiments, we found that the 606 cm(-)(1) mode is upshifted to about 618 cm(-)(1) by Sr(2+) substitution but that this mode is not affected by substitution with the (44)Ca isotope. On the basis of these results and also on the basis of studies of Mn model compounds, we assign the 625 cm(-)(1) mode in the S(1) state and the 606 cm(-)(1) mode in the S(2) state to a Mn-O-Mn cluster vibration of the oxygen-evolving complex (OEC) in PSII. This structure may include additional bridge(s), which could be another oxo, carboxylato(s), or atoms derived from an amino acid side chain. Our results indicate that the bridged oxygen atom shown in this Mn-O-Mn cluster is exchangeable and accessible by water. The downshift in the Mn-O-Mn cluster vibration as manganese is oxidized during the S(1) --> S(2) transition is counterintuitive; we discuss possible origins of this behavior. Our results also indicate that Sr(2+) substitution in PSII causes a small structural perturbation that affects the bond strength of the Mn-O-Mn cluster in the PSII OEC. This suggests that Sr(2+), and by inference, Ca(2+), communicates with, but is not integral to, the manganese core.     

The effects of dissolved oxygen levels on the metabolic interaction between digestion and locomotion in Cyprinid fishes with different locomotive and digestive performances

To test whether the effects of water oxygen concentration ([O(2)]) on the metabolic interaction between locomotion and digestion differ between fish species with different locomotive and digestive behaviours in normoxia, we investigated the swimming performance of fasted and fed fish at water [O(2)] of 1, 2 and 8 (normoxia) mg L(-1) (2.5, 5 and 20 kPa) at 25°C in three juvenile Cyprinidae fish species: goldfish (Carassius auratus), common carp (Cyprinus carpio) and qingbo (Spinibarbus sinensis). Digestion, taxon and water [O(2)] all had significant effects on the pre-exercise oxygen consumption rate [Formula: see text] and the swimming performance (P < 0.05). Among the three fishes, qingbo showed the highest swimming performance and the lowest feeding [Formula: see text] at the saturated water [O(2)], and its active oxygen consumption rate [Formula: see text] and critical swimming speed (U (crit)) decreased the most with decreases in water [O(2)]. Qingbo exhibited a locomotion-priority metabolic mode at all three water [O(2)]. Digestion was sacrificed to locomotion in a postprandial swimming situation, but fed qingbo could not maintain their U (crit) at water [O(2)] of 2 and 1 mg L(-1). Goldfish showed the lowest swimming performance and the highest feeding [Formula: see text] at the saturated water [O(2)]. They exhibited a digestion-priority metabolic mode at high water [O(2)]. However, with a decrease in water [O(2)], the feeding [Formula: see text] decreased more acutely than the respiratory capacity; thus, digestion and locomotion performed independently in a postprandial swimming situation (i.e., an additive metabolic mode) at a water [O(2)] of 1 mg L(-1). The common carp showed moderate and balanced swimming performance and feeding [Formula: see text] at the saturated water [O(2)], and exhibited an additive metabolic mode at all 3 water [O(2)], because digestion, swimming and respiratory capacities decreased in parallel with the decrease in water [O(2)].     

Protein unfolding in detergents: effect of micelle structure,ionic strength,pH, and temperature

The 101-residue monomeric protein S6 unfolds in the anionic detergent sodium dodecyl sulfate (SDS) above the critical micelle concentration, with unfolding rates varying according to two different modes. Our group has proposed that spherical micelles lead to saturation kinetics in unfolding (mode 1), while cylindrical micelles prevalent at higher SDS concentrations induce a power-law dependent increase in the unfolding rate (mode 2). Here I investigate in more detail how micellar properties affect protein unfolding. High NaCl concentrations, which induce cylindrical micelles, favor mode 2. This is consistent with our model, though other effects such as electrostatic screening cannot be discounted. Furthermore, unfolding does not occur in mode 2 in the cationic detergent LTAB, which is unable to form cylindrical micelles. A strong retardation of unfolding occurs at higher LTAB concentrations, possibly due to the formation of dead-end protein-detergent complexes. A similar, albeit much weaker, effect is seen in SDS in the absence of salt. Chymotrypsin inhibitor 2 exhibits the same modes of unfolding in SDS as S6, indicating that this type of protein unfolding is not specific for S6. The unfolding process in mode 1 has an activation barrier similar in magnitude to that in water, while the activation barrier in mode 2 is strongly concentration-dependent. The strong pH-dependence of unfolding in SDS and LTAB suggests that the rate of unfolding in anionic detergent is modulated by repulsion between detergent headgroups and anionic side chains, while cationic side chains modulate unfolding rates in cationic detergents.     

不同养殖模式对亚东鲑幼鱼生长性能、血清生化指标和肌肉营养成分的影响

为了探索亚东鲑(Salmon trutta)多元化养殖模式,以初始体质量(100.05±1.12) g幼鱼作为研究对象,开展循环水和开放流水模式养殖比较,试验周期180d,分析比较其生长性能、血清生化指标和肌肉营养成分差异。结果表明:在开放流水模式下增重率、肥满度、肝体指数、特定生长率和饲料转化率均显著高于循环水模式(P<0.05),存活率二者差异不显著(P>0.05);在循环水模式下补体C₄、总蛋白和生长激素均显著低于开放流水模式(P<0.05),但溶菌酶、谷丙转氨酶和谷草转氨酶相反(P<0.05),补体C₃、碱性磷酸酶、酸性磷酸酶和球蛋白在两种模式之间差异均不显著(P>0.05);肌肉主要营养成分粗蛋白、粗脂肪含量开放流水模式高于循环水模式,氨基酸组成二者差异不显著(P>0.05),脂肪酸中油酸、α-亚麻酸、C20:1、C20:2和MUFA含量均显著低于开放流水(P<0.05),而EPA+DHA、SAFA和PUFA含量相反,二者之间差异性显著(P<0.05),矿物质镁含量循环水模式显著高于开...     

Water filtration rate and infiltration/accumulation of low density lipoproteins in 3 different modes of endothelial/smooth muscle cell co-cultures

Using different endothelial/smooth muscle cell co-culture modes to simulate the intimal structure of blood vessels, the water filtration rate and the infiltration/accumulation of LDL of the cultured cell layers were studied. The three cell culture modes of the study were: (i) The endothelial cell monolayer (EC/Φ); (ii) endothelial cells directly co-cultured on the smooth muscle cell monolayer (EC-SMC); (iii) endothelial cells and smooth muscle cells cultured on different sides of a Millicell-CM membrane (EC/SMC). It was found that under the same condition, the water filtration rate was the lowest for the EC/SMC mode and the highest for the EC/Φ mode, while the infiltration/accumulation of DiI-LDLs was the lowest in the EC/Φ mode and the highest in the EC-SMC mode. It was also found that DiI-LDL infiltration/accumulation in the cultured cell layers increased with the increasing water filtration rate. The results from the in vitro model study therefore suggest that the infiltration/accumulation of the lipids within the arterial wall is positively correlated with concentration polarization of atherogenic lipids, and the integrity of the endothelium plays an important role in the penetration and accumulation of atherogenic lipids in blood vessel walls.     

Interactions of water soluble porphyrins with Z-poly(dG-dC).

The water soluble porphyrin tetrakis(4-N-methylpyridyl)porphine (H2TMpyP) and its copper(II) derivative (CuTMpyP) convert Z-poly(dG-dC) to the B-form. For H2TMpyP, the fraction Z character (fr-Z) is given by fr-Z = 1.0 - 21 rO and for CuTMpyP, fr-Z = .94 - 12 rO where rO identical to [Porphyrin]O/[DNA]O. Neither the manganese(III) derivative of of this porphyrin (MnTMpyP) nor tetrakis(2-N-methylpyridyl)porphine (H2TMpyP-2) is nearly as effective at causing the conversion. The former two porphyrins have been shown to intercalate into B-poly(dG-dC) whereas the latter two porphyrins do not. The kinetics of the Z----B conversion are independent of porphyrin or poly(dG-dC) concentration for 1/rO greater than 6. At smaller values of 1/rO, the conversion rate is greatly increased for H2TMpyP and CuTMpyP. The interaction of these porphyrins with Z-poly(dG-dC) follows simple first order kinetics in this latter concentration range. It is proposed that for small values of 1/rO the sequence of events begins with a porphyrin-unassisted distortion of the Z-duplex (with a rate constant of 0.6 s-1) followed by a rapid uptake of porphyrin in what may be an intercalative mode. The porphyrin thus located in Z-regions brings about rapid conversion to the B-form. Binding of H2TMpyP or CuTMpyP to B-regions of a predominantly Z-strand leads to conversion of Z to B. However, this conversion process is considerably slower than when the porphyrins bind directly to Z-regions.     

The basis for k(cat) impairment in prophospholipase A(2) from the anion-assisted dimer structure

Kinetic results in this paper show that, contrary to earlier reports, pig pancreatic prophospholipase A(2) (proPLA2) does not hydrolyze monodisperse short chain phosphatidylcholine below the critical micelle concentration. ProPLA2 is active on an anionic interface, but at a rate that is decreased by more than 100-fold compared to that of PLA2, the active form. Solution studies show that both proPLA2 and PLA2 bind to an anionic interface and also bind a tetrahedral intermediate mimic at the active site. The 1.5 A resolution crystal structure of the anion-assisted dimer of proPLA2 reported in this paper is compared with the corresponding structure for PLA2 [Pan, Y. H., et al. (2001) Biochemistry 40, 609-617]. As a mimic for the forms bound to the anionic interface, these structures provide insights into the possible structural basis for the impaired chemical step of the zymogen. The proPLA2 dimer contained within one crystallographic asymmetric unit has one molecule of the inhibitor 1-hexadecyl-3-(trifluoroethyl)-sn-glycero-2-phosphomethanol and is bridged by four coplanar sulfate anions. Relative to the structure of PLA2, the subunit contact surface in proPLA2 displays a tilted orientation, an altered mode of inhibitor binding, displacement of a mechanistically significant loop that includes Tyr69, and a critical active site water seen in PLA2 that is not seen in proPLA2. These differences are interpreted to suggest possible origins of the functional differences between the pro and active enzyme at an anionic interface. A structural origin of this difference is discussed in terms of the calcium-coordinated activated water mechanism of the esterolysis reaction. Together, a comparison of the structures of the anion-assisted dimers of PLA2 and proPLA2 not only offers an explanation of why the zymogen form is k(cat)-impaired and binds poorly even to the anionic interface but also supports a mechanism for the activated enzyme that includes a critical second-sphere assisting water bridging His48 and the calcium-coordinated catalytic water.     

Sexual and reproductive traits of Hypania invalida (Polychaeta,Ampharetidae): a remarkable invasive species in Central European waterways

1. The Ponto‐Caspian polychaete Hypania invalida (Archiv für Naturgeschichte, 1860, 26, 109) is undergoing rapid range expansion in the Rhine and other central European waterways. We examined its reproductive traits in an attempt to account for its remarkable invasive success. 2. For the first time in this species, we found males, dioecy (gonochorism) and an exclusively sexual mode of reproduction; no indication for hermaphroditism or (unisexual) parthenogenesis, that could explain the rapid range expansion of H. invalida, was found. 3. Our experimental evidence shows that H. invalida reproduces by males discharging their sperm into the water column while eggs are retained and fertilised in the female dwelling tubes. This mode of ‘spermcast’ reproduction is common in sessile benthic animals on wave‐washed shores. 4. Fecundity of H. invalida ranged between 60 and 260 eggs per clutch and is increased by iteroparity (i.e. multiple cycles of oogenesis and egg‐laying per specimen). Offspring are brooded for about 2 weeks before they leave the maternal dwelling tubes and disperse in the water column. 5. After connection to a supply of Rhine water, sediments in flumes were rapidly colonised up to a density of 12 000 ind. m^?2 via the settlement of small larvae (<300 μm) from the water column; these reached sexual maturity approximately 12 weeks after settlement. 6. The reproductive traits of H. invalida give the species a high rate of dispersal. The capacity of its larvae to survive passage through pumps before settlement enables them to travel in the ballast water of ships. This might explain the upstream expansion of H. invalida in rivers. We conclude that further range expansion, and especially intercontinental transport of this polychaete, can only be prevented by improved ballast water treatment.     

Synthesis and structure-activity relationship of 4-(1,3-benzothiazol-2-yl)-thiophene-2-sulfonamides as cyclin-dependent kinase 5 (cdk5)/p25 inhibitors

4-(1,3-Benzothiazol-2-yl)thiophene-2-sulfonamide (4a) was found to be a moderately potent inhibitor of cyclin-dependent kinase 5 (cdk5) from a HTS screen. The synthesis and SAR around this hit is described. The X-ray coordinates of ligand 4a with cdk5 are also reported, showing an unusual binding mode to the hinge region via a water molecule.     

Kinetic fluorescence measurement of fluorescein di-beta-D-galactoside hydrolysis by beta-galactosidase: intermediate channeling in stepwise catalysis by a free single enzyme

Kinetic fluorescence measurements were employed to quantitative to stepwise hydrolysis of fluorescein di-beta-D-galactoside (FDG) by beta-galactosidase and the intermediate fluorescein mono-beta-D-galactoside (FMG) channeling. The kinetic parameters, Michaelis-Menten constant Km and enzymatic catalysis rate k2, for FDG hydrolysis to FMG by beta-galactosidase were obtained as 18.0 microM and 1.9 mumol.(min-mg)-1, respectively. The FMG intermediate is hydrolyzed via two modes: (1) FMG that is in free solution binding to the enzyme substrate binding site in competition with FDG and then being hydrolyzed (binding mode); (2) FMG being directly hydrolyzed into the final products of fluorescein and galactose before the FMG can diffuse away from the enzyme active site (channeling mode). The extent of the FMG channeling mode was found to depend on the FDG hydrolysis rate but to be independent of the free enzyme concentration. A channeling factor, defined as the ratio of the real FMG hydrolysis rate with both binding and channeling modes over that which would be observed with an exclusive binding mode, was used to quantitate the effect of the intermediate channeling. The FMG channeling factor was determined to be close to 1 at low FDG concentration (about 5.1 microM), where the slow FDG hydrolysis rate gives an ineffective channeling and where the FMG is then hydrolyzed mainly via the binding mode. However, the channeling factor dramatically increases at higher FDG concentrations (greater than Km), strongly indicating that the effective FMG channeling mode, resulting from the considerable FDG hydrolysis rate at high FDG concentrations, becomes a primary pathway to channel a steady system hydrolysis with a high rate.(ABSTRACT TRUNCATED AT 250 WORDS)     

Disinfection of drinking water by using a novel electrochemical reactor employing carbon-cloth electrodes.

A novel electrochemical reactor employing carbon-cloth electrodes was constructed for disinfection of drinking water. Escherichia coli K-12 (10(2) cells per cm3) was sterilized when a cell suspension was passed through the reactor at a dilution rate of 6.0 h-1, and a potential of 0.7 V versus a saturated calomel electrode was applied to an electrode. The survival ratio increased with increasing dilution rate but was less than 0.1% at dilution rates of less than 6.0 h-1. Although the survival ratio increased with increasing cell concentration above 10(3) cells per cm3, the disinfection rate also increased. The disinfection rate was 6.0 x 10(2) cells per cm3 per h at a cell concentration of 10(2) cells per cm3. Continuous sterilization of E. coli cells was carried out for 24 h. Sterilization is based on an electrochemical reaction between the electrode and the cell which is mediated by intracellular coenzyme A. Sterilization of drinking water by using this reactor was successfully performed, demonstrating the potential of such a reactor for clean and efficient water purification.     

Origin of the anomalous Fe-CO stretching mode in the CO complex of Ascaris hemoglobin

We report an unusually high frequency (543 cm(-)(1)) for an Fe-CO stretching mode in the CO complex of Ascaris suum hemoglobin as compared to vertebrate hemoglobins in which the frequency of the Fe-CO mode is much lower. A second Fe-CO stretching mode in Ascaris hemoglobin is observed at 515 cm(-1). We propose that these two Fe-CO stretching modes arise from two protein conformers corresponding to interactions of the heme-bound CO with the B10-tyrosine or the E7-glutamine residues. This postulate is supported by spectra from the B10-Tyr --> Phe mutant in which the 543 cm(-1) line is absent. Thus, a strong polar interaction, such as hydrogen bonding, of the CO with the distal B10 tyrosine residue is the dominant factor that causes this anomalously high frequency. Strong hydrogen bonding between O(2) and distal residues in the oxy complex of Ascaris hemoglobin has been shown to result in a rigid structure, rendering an extremely low oxygen off rate [Gibson, Q. H., and Smith, M. H. (1965) Proc. R. Soc. London B 163, 206-214]. In contrast, the CO off rate in Ascaris hemoglobin is very similar to that in sperm whale myoglobin. The high CO off rate relative to that of O(2) in Ascaris hemoglobin is attributed to a rapid equilibrium between the two conformations of the protein in the CO adduct, with the off rate being determined by the conformer with the higher rate.     

河西走廊绿洲灌区循环模式"农田-食用菌"生产系统氮素流动特征

河西走廊绿洲灌区在农业产业化进程加快的同时,也带来了水资源和生态环境的压力.本文利用养分流动和模型分析的方法,分析河西走廊绿洲灌区典型“农田-食用菌”生产系统(农田-食用菌集约生产模式、农田-食用菌单户生产模式、农田单作)的氮元素流动特征.结果表明:农田作物秸秆通过食用菌体系还田使氮素利用率提高了10％左右,秸秆还田氮输入量(165.6kg· hm-2·a-1)占农田氮素总输入量的37.1％,使化肥氮输入量减少,因此秸秆氮的合理循环利用可作为减少化肥投入的有效途径.但农田氮素仍有盈余,单位面积氮盈余量高达217.0 kg·hm-2· a-1,未能实现循环模式内养分平衡的理想效果,因此优化氮素管理、确定合理的大田作物和食用菌面积、调整农业产业结构是解决该问题的关键.     

Disinfection of drinking water by using a novel electrochemical reactor employing carbon-cloth electrodes.

A novel electrochemical reactor employing carbon-cloth electrodes was constructed for disinfection of drinking water. Escherichia coli K-12 (10(2) cells per cm3) was sterilized when a cell suspension was passed through the reactor at a dilution rate of 6.0 h-1, and a potential of 0.7 V versus a saturated calomel electrode was applied to an electrode. The survival ratio increased with increasing dilution rate but was less than 0.1% at dilution rates of less than 6.0 h-1. Although the survival ratio increased with increasing cell concentration above 10(3) cells per cm3, the disinfection rate also increased. The disinfection rate was 6.0 x 10(2) cells per cm3 per h at a cell concentration of 10(2) cells per cm3. Continuous sterilization of E. coli cells was carried out for 24 h. Sterilization is based on an electrochemical reaction between the electrode and the cell which is mediated by intracellular coenzyme A. Sterilization of drinking water by using this reactor was successfully performed, demonstrating the potential of such a reactor for clean and efficient water purification.     

Water Balance in Cucumis Plants, Measured by Nuclear Magnetic Resonance, II

Nuclear magnetic resonance (NMR) was used to investigate theeffects of changes in root temperature, of changes in the areaof root in contact with culture solution and of day/night rhythmon the water balance of a cucumber and a gherkin plant. Resultsare discussed in terms of water potential, flow rate and resistanceusing a previously presented model of water balance. As longas water uptake alone is varied, flow rate and water content(or potential) will change in the same direction. In contrast,from that model it is predicted that changes in transpirationwill affect flow rate and water content in opposite ways. Anexperimental verification of this prediction was given in theprevious paper. Results obtained by the NMR method are comparedto those determined using a dendrometer. The results demonstratethat the NMR method is a valuable tool to study plant waterbalance and that it can serve as a technique for discriminatingbetween changes in plant water balance that are due to changesin water uptake by roots and those due to changes in transpiration. Key words: Water balance model, Cucumis satious L., flow, water content, NMR, water balance measurement     

Excess postexercise oxygen consumption is unaffected by the resistance and aerobic exercise order in an exercise session

The main purpose of this study was to compare the magnitude and duration of excess postexercise oxygen consumption (EPOC) after 2 exercise sessions with different exercise mode orders, resistance followed by aerobic exercise (R-A); aerobic by resistance exercise (A-R). Seven young men (19.6 ± 1.4 years) randomly underwent the 2 sessions. Aerobic exercise was performed on a treadmill for 30 minutes (80-85% of reserve heart rate). Resistance exercise consisted of 3 sets of 10 repetition maximum on 5 exercises. Previous to the exercise sessions, V(O2), heart rate, V(CO2), and respiratory exchange rate (RER) were measured for 15 minutes and again during recovery from exercise for 60 minutes. The EPOC magnitude was not significantly different between R-A (5.17 ± 2.26 L) and A-R (5.23 ± 2.48 L). Throughout the recovery period (60 minutes), V(O2) and HR values were significantly higher than those observed in the pre-exercise period (p < 0.05) in both exercise sessions. In the first 10 minutes of recovery, V(CO2) and RER declined to pre-exercise levels. Moreover, V(CO2) and RER values in A-R were significantly lower than in R-A. In conclusion, the main result of this study suggests that exercise mode order does not affect the EPOC magnitude and duration. Therefore, it is not necessary for an individual to consider the EPOC when making the decision as to which exercise mode is better to start a training session.     

Coupling of electron and proton transfer in oxidative water cleavage in photosynthesis

This minireview addresses questions on the mechanism of oxidative water cleavage with special emphasis on the coupling of electron (ET) and proton transfer (PT) of each individual redox step of the reaction sequence and on the mode of O-O bond formation. The following topics are discussed: (1) the multiphasic kinetics of Y(Z)(ox) formation by P680(+*) originate from three different types of rate limitations: (i) nonadiabatic electron transfer for the "fast" ns reaction, (ii) local "dielectric" relaxation for the "slow" ns reaction, and (iii) "large-scale" proton shift for the micros kinetics; (2) the ET/PT-coupling mode of the individual redox transitions within the water oxidizing complex (WOC) driven by Y(Z)(ox) is assumed to depend on the redox state S(i): the oxidation steps of S(0) and S(1) comprise separate ET and PT pathways while those of S(2) and S(3) take place via proton-coupled electron transfer (PCET) analogous to Jerry Babcock's hydrogen atom abstractor model [Biochim. Biophys. Acta, 1458 (2000) 199]; (3) S(3) is postulated to be a multistate redox level of the WOC with fast dynamic equilibria of both redox isomerism and proton tautomerism. The primary event in the essential O-O bond formation is the population of a state S(3)(P) characterized by an electronic configuration and nuclear geometry that corresponds with a complexed hydrogen peroxide; (4) the peroxidic type S(3)(P) is the entatic state for formation of complexed molecular oxygen through S(3) oxidation by Y(Z)(ox); and (5) the protein matrix itself is proposed to exert catalytic activity by functioning as "PCET director". The WOC is envisaged as a supermolecule that is especially tailored for oxidative water cleavage and acts as a molecular machine.     

An undiscovered facet of hydraulic redistribution driven by evaporation—a study from a Populus tomentosa plantation

Maintaining the activity and function of the shallow root system of plants is essential for withstanding drought stress, but the associated mechanism is poorly understood. By investigating sap flow in 14 lateral roots (LRs) randomly selected from trees of a Chinese white poplar (Populus tomentosa) plantation receiving three levels of irrigation, an unknown root water transport mode of simultaneous daytime bi-directional water flow was discovered. This mode existed in five LRs confined to the surface soil without attached sinker roots. In the longer term, the bi-directional water flow was correlated with the soil water content. However, within the day, it was associated with transpiration. Our data demonstrated that bi-directional root sap flow occurred during the day, and was driven by evaporative demand, further suggesting the existence of circumferential water movement in the LR xylem. We named this phenomenon evaporation-driven hydraulic redistribution (EDHR). A soil-root water transport model was proposed to encapsulate this water movement mode. EDHR may be a crucial drought-tolerance mechanism that allows plants to maintain shallow root survival and activity by promoting root water recharge under extremely dry conditions.     

涌泉根灌下水氮耦合对陕北山地苹果光合特性、产量和水氮利用的影响

以陕北山地7年生‘寒富’苹果树为试验材料,设置3个灌水水平[高水(W₁,85%～100%θ_f,θ_f为田间持水量)、中水(W₂,70%～85%θ_f)、低水(W₃,55%～70%θ_f)]和3个施氮水平[高氮(N₁,600 kg·hm^-2)、中氮(N₂,400 kg·hm^-2)、低氮(N₃,200 kg·hm^-2)],研究涌泉根灌条件下水氮耦合对山地苹果树光合特性、产量和水氮利用的影响。结果表明: 相同灌水条件下,随着施氮量的减少,苹果树叶片的净光合速率(P_n)、蒸腾速率(T_r)、气孔导度(g_s)和瞬时水分利用效率(WUE_i)降低,但胞间CO₂浓度(C_i)增加;相同施氮条件下,随着灌水量的减少,叶片P_n、T_r、g_s和WUE_i降低,而C_i增加。W₁N₁处理的P_n、T_r日均值最大,但与W₁N₁处理相比,W₂N₂处理的WUE_i最大。苹果产量、灌溉水利用效率(IWUE)和氮肥偏生产力(NPFP)受灌水和施氮量的显著影响,W₂N₂处理的产量最高(26761 kg·hm^-2),减小灌水量和增大施氮量使IWUE显著提高,而增大灌水量和降低施氮量使NPFP显著增加。回归分析表明,产量和IWUE同时获得最优解时,灌水量和施氮量组合最接近W₂N₂处理。因此,W₂N₂处理为涌泉根灌条件下陕北山地苹果最佳的水氮组合模式。     

Interfacial catalysis by phospholipase A2: evaluation of the interfacial rate constants by steady-state isotope effect studies.

The kinetics of hydrolysis of phospholipid vesicles by phospholipase A2 (PLA2) in the scooting mode can be described by the Michaelis-Menten formalism for the action of the enzyme in the interface (E*). E* + S in equilibrium E*S in equilibrium E*P in equilibrium E* + Products The values of the interfacial rate constants cannot be obtained by classical methods because the concentration of the substrate within the lipid bilayer is not easily manipulated. In the present study, carbonyl-carbon heavy atom isotope effects for the hydrolysis of phospholipids have been measured in both vesicles and in mixed micelles in which the phospholipid was present in the nonionic detergent Triton X-100. A large [14C]carbonyl carbon isotope effect of 1.12 +/- 0.02 was measured for the cobra venom PLA2-catalyzed hydrolysis of dipalmitoylphosphatidylcholine in Triton X-100. In contrast, no isotope effect (1.01 +/- 0.01) was measured for the action of the porcine pancreatic and cobra venom enzymes on vesicles of dimyristoylphosphatidylmethanol in the scooting mode. In a second experiment, the hydrolysis of vesicles was carried out in oxygen-18 enriched water. Analysis of the released fatty acid product by mass spectrometry showed that it contained only a single oxygen-18. All of these results were used to estimate both the forward and reverse commitments to catalysis. The lack of doubly labeled fatty acid demonstrated that the product is released from the E*P complex faster than the reverse of the esterolysis step. The small isotope effect in vesicles demonstrated that the E*S complex goes on to products faster than substrate is released from the enzyme. The relevance of these results to an understanding of substrate specificity and inhibition of PLA2 is discussed. In addition, the conditions placed on the values of the rate constants obtained in the present study together with results obtained in the other studies described in this series of papers have led to the evaluation of most of the interfacial rate constants for the hydrolysis of phospholipid vesicles by PLA2.