大针茅典型草原土壤CO2排放规律的研究

采用静态室/碱吸收法研究了内蒙古大针茅典型草原土壤呼吸的时空动态.大针茅群落地上部和地下部生物量的时空变异不同步,用地上部生物量不能很好地预测群落的根系量.群落土壤呼吸具有明显的时空动态,控制群落土壤呼吸空间变异与时间变异的因子是不同的.土壤呼吸的季节变化与地上部生物量及土壤水分状况关系密切.根据大针茅群落分别计算了大针茅群落1995、1997和1998年的年CO₂排放总量,分别为180、45.8和225gC·m^-2·a^-1,年际变异很大.过度放牧大大降低了群落的生物量,也降低了土壤CO₂排放量.讨论了建立以降水量为驱动因子的草地群落土壤呼吸动态模型的可能性.     

环境因子和生物因子对黄河三角洲滨海湿地土壤呼吸的影响

采用Li-8150多通道土壤呼吸自动测量系统对黄河三角洲滨海湿地土壤呼吸进行全年连续测定,同步测量了温度、土壤含水量、地上生物量以及叶面积指数等环境因子和生物因子.结果表明: 土壤呼吸日动态在全年尺度上多呈单峰型,但在受到土壤封冻和地表积水干扰时,土壤呼吸日动态呈多峰型.土壤呼吸具有明显的季节动态特征,总体呈单峰型,年平均土壤呼吸速率为0.85 μmol CO₂·m^-2·s^-1,生长季平均土壤呼吸速率为1.22 μmol CO₂·m^-2·s^-1.在全年尺度上,土壤温度是滨海湿地土壤呼吸的主要控制因子,可解释全年土壤呼吸87.5%的变化.在生长季尺度上,土壤含水量和叶面积指数对土壤呼吸的协同影响达到85%.     

水稻光合碳在植物-土壤系统中的分配及其对CO2升高和施氮的响应

采用¹³C-CO₂进行连续标记,研究水稻分蘖期和孕穗期光合碳在植株-土壤系统中的分配及其对大气CO₂浓度升高(800 μL·L^-1)和施氮(100 mg·kg^-1)的响应.结果表明: CO₂浓度升高显著提高分蘖期根系生物量和孕穗期地上部生物量,并使生物量根冠比在分蘖期增加,而在孕穗期减小.CO₂浓度升高条件下,施氮使水稻地上部分生物量增加,却显著降低了孕穗期水稻根系生物量.CO₂浓度升高使光合¹³C在孕穗期向土壤的输入显著增加,然而施肥并没有促进由CO₂浓度升高驱动的光合¹³C在土壤中的积累,而且还降低了土壤中的光合¹³C的分配比例.综上,CO₂浓度升高显著提高了稻田土壤光合碳输入,促进稻田有机碳周转;施氮促进了水稻地上部的生长,却降低了光合碳向地下的分配比例.     

水稻光合碳在植物-土壤系统中的分配及其对CO2升高和施氮的响应

采用¹³C-CO₂进行连续标记,研究水稻分蘖期和孕穗期光合碳在植株-土壤系统中的分配及其对大气CO₂浓度升高(800 μL·L^-1)和施氮(100 mg·kg^-1)的响应.结果表明: CO₂浓度升高显著提高分蘖期根系生物量和孕穗期地上部生物量,并使生物量根冠比在分蘖期增加,而在孕穗期减小.CO₂浓度升高条件下,施氮使水稻地上部分生物量增加,却显著降低了孕穗期水稻根系生物量.CO₂浓度升高使光合¹³C在孕穗期向土壤的输入显著增加,然而施肥并没有促进由CO₂浓度升高驱动的光合¹³C在土壤中的积累,而且还降低了土壤中的光合¹³C的分配比例.综上,CO₂浓度升高显著提高了稻田土壤光合碳输入,促进稻田有机碳周转;施氮促进了水稻地上部的生长,却降低了光合碳向地下的分配比例.     

短期增温和增加降水对内蒙古荒漠草原土壤呼吸的影响

利用红外辐射增温装置模拟短期持续增温和降水增加交互作用对内蒙古荒漠草原土壤呼吸作用的影响, 结果表明: 土壤含水量对月土壤呼吸的影响显著大于土壤温度增加的影响, 生长旺季的月土壤呼吸显著大于生长末季; 土壤温度和水分增加都显著影响日土壤呼吸, 但二者的交互作用对土壤呼吸无显著影响。荒漠草原7‒8月平均土壤呼吸速率为1.35 μmol CO₂·m ^-2·s ^-1, 7月份为2.08 μmol CO₂·m ^-2·s ^-1, 8月份为0.63 μmol CO₂·m ^-2·s ^-1。土壤呼吸与地下各层根系生物量呈幂函数关系, 0‒10 cm土层的根系生物量对土壤呼吸的解释率(79.2%)明显高于10‒20 cm土层的解释率(31.6%)。0-10 cm土层的根系生物量是根系生物量的主体, 根系生物量对土壤呼吸的影响具有层次性。在未来全球变暖和降水格局变化的情景下, 荒漠草原土壤水分含量是影响生物量的主导环境因子, 而根系生物量的差异是造成土壤呼吸异质性的主要生物因素, 土壤含水量可通过影响根系生物量控制土壤呼吸的异质性。     

川中丘陵区水稻田土壤呼吸及其影响因素

基于川中丘陵区2003年4~9月水稻田土壤呼吸、土壤温度和水稻(Oryza sativa)生物量的测定,研究了水稻田土壤呼吸日变化和季节变化特征以及影响稻田土壤呼吸的主要因素。结果表明,水稻田土壤CO₂排放通量的日变化为单峰型,其最小值和最大值分别出现在当地时间7∶00和15∶00;在水稻生长期内,稻田土壤CO₂排放通量在18.00~269.69 mg·m^-2·h^-1之间波动,平均排放通量为121.76 mg·m^-2·h^-1。在日的时间尺度上,水稻田土壤CO₂排放通量与5 cm土壤温度存在显著的指数函数关系;而从整个生长期时间尺度上看,水稻田土壤CO₂的排放通量主要受到5 cm土壤温度和水稻地下生物量的影响。在水稻生长初期,水稻地下生物量与稻田土壤CO₂排放通量之间存在着显著的相关关系;水稻拔节中后期到成熟期,土壤温度则是制约稻田土壤CO₂排放的关键因素。CO₂排放通量与稻田地表水层深度的相关关系不显著。     

渭河平原农田冬小麦土壤呼吸及其影响因素

基于对半湿润易旱区的渭河平原农田2013—2014年冬小麦生长期土壤呼吸(SR)及环境因子和生物因子的观测,研究了冬小麦土壤呼吸日变化、季节变化特征,综合分析了温度(T)、土壤含水量(W)、总初级生产力(GPP)和叶面积指数(LAI)对土壤呼吸的影响.结果表明: 冬小麦土壤呼吸日变化呈单峰型,呼吸速率变化范围为1.5～6.94 μmol CO₂·m^-2·s^-1,最大值出现在12:00—14:00;温度是影响土壤呼吸日变化的驱动因子,其中地表温度(T_s)能解释土壤呼吸时间变异的80.9%;土壤呼吸速率与温度的昼夜变化对应关系呈顺时针近椭圆曲线.冬小麦土壤呼吸速率从出苗后到冬季呈下降趋势,在冬季时保持较低水平,进入返青期后迅速增加,在抽穗期和灌浆期达到最大,成熟期后有所下降,变化范围为0.65～4.85 μmol CO₂·m^-2·s^-1;土壤呼吸季节变化与温度、土壤含水量、GPP、LAI均呈显著(P<0.01)正相关关系;土壤温度和水分是影响土壤呼吸季节变化的关键因素,使用复合模型SR=e^{(a+bT_{5 cm}}+cW10 cm+dW_{10 cm²}),可以解释土壤呼吸时间变异的82.6%,比单因子模型(不超过65.7%)的解释能力显著提高.经模型计算,该区域2013—2014年冬小麦生长期平均土壤呼吸速率为1.67 μmol CO₂·m^-2·s^-1.     

玉米地土壤呼吸作用对土壤温度和生物因子协同作用的响应

基于2005年玉米(Zea mays)生长季土壤呼吸作用及其影响因子的动态观测资料,分析了玉米地土壤呼吸作用的日和季动态及其对土壤温度和生物因子协同作用的响应。结果表明,玉米地土壤呼吸作用的日变化为不对称的单峰型,其最小值和最大值分别出现在6∶00~7∶00和13∶00左右;玉米生长季中,土壤呼吸速率波动较大,其均值为3.16 μmol CO₂·m^-2·s^-1,最大值为4.87 μmol CO₂·m^-2·s^-1,出现在7月28日,最小值为1.32 μmol CO₂·m^-2·s^-1,出现在5月4日。在土壤呼吸作用日变化中,土壤呼吸速率(SR)与10 cm深度土壤温度(T)呈显著的线性关系:SR=αT+β。在整个生长季节,玉米净初级生产力(NPP)与直线斜率(α)呈显著正相关,生物量(B)也明显影响直线的截距(β)。基于此,建立了玉米地土壤呼吸作用动态模型SR=(aNPP+b)T+cB²+dB+e。土壤呼吸作用季节变化的大部分(97%)可以由土壤温度、NPP和生物量的季节变化来解释。当仅考虑土壤温度对土壤呼吸作用的影响时,指数方程会过大或过小地估计了土壤呼吸强度。该文的结果强调了生物因子在土壤呼吸作用季节变化中的重要作用,同时指出土壤呼吸作用模型不仅要考虑土壤温度的影响,在生物因子影响土壤呼吸作用的温度敏感性时,还应该把生物因子纳入模型。     

呼伦贝尔地区贝加尔针茅草甸草原土壤CO2排放特征

采用Li-8150多通道土壤碳通量自动观测系统,于2009年6月—2010年6月对呼伦贝尔地区贝加尔针茅草甸草原土壤呼吸进行连续野外观测,分析该地区土壤CO₂通量排放特征.结果表明: 生长季内贝加尔针茅草甸草原土壤呼吸日动态呈单峰曲线,最高值出现在13:00—15:00,最低值出现在5:00—6:00,土壤呼吸呈明显的季节变化,与土壤温度和土壤含水量季节动态相吻合.土壤呼吸与各层土壤温度和土壤含水量关系可以用线性模型和指数-乘幂模型来表示;土壤呼吸与各层土壤温度呈显著的指数回归关系,Q₁₀变化范围分别为1.68～2.14和3.03～3.60,非生长季土壤温度对土壤呼吸的影响更为显著;生长季内土壤呼吸与10 cm土壤含水量呈显著正相关.2009和2010年土壤CO₂年排放量分别为488.47和507.20 g C·m^-2·a^-1,生长季排放量约占年排放量的90%.     

松嫩平原西部草甸草原典型植物群落土壤呼吸动态及影响因素

以松嫩平原西部草甸草原中典型植物虎尾草、碱茅、芦苇和羊草群落为对象,分析了4种植被群落土壤呼吸速率日动态和季节动态及其影响因素,以及土壤盐碱度与土壤呼吸碳排放量的关系.结果表明： 4种植物群落的土壤呼吸速率日变化均呈明显的单峰曲线,峰值出现在11:00-15:00,而谷值大多出现在21:00-1:00或3:00-5:00;4种植被群落土壤呼吸速率的季节变化趋势一致,7、8月的土壤呼吸速率(3.21~4.84 μmol CO₂·m^-2·s^-1)最高,10月最低(0.46~1.51 μmol CO₂·m^-2·s^-1);各群落土壤呼吸速率与土壤和近地表大气温度之间呈极显著相关关系,其中,虎尾草群落的土壤呼吸速率与土壤表层含水量极显著相关,芦苇和羊草群落土壤呼吸速率与近地表的相对湿度显著相关.土壤盐分含量明显抑制了土壤CO₂排放量,土壤pH、电导率和土壤交换性钠可以解释该草甸草原土壤呼吸空间变异的87%～91%.     

ATP/ADP exchange activity of gastric (H+ + K+)-ATPase

The ATP/ADP exchange is shown to be a partial reaction of the $\text{(}\text{H}{}^{\mathrm{+}}\text{+}\text{K}{}^{\mathrm{+}}\text{)-}\text{ATPase}$ by the absence of measurable nucleoside diphosphokinase activity and the insensitivity of the reaction to $\text{P}{}^1$, $\text{P}{}^5$ -di(adenosine-5′) pentaphosphate, a myokinase inhibitor. The exchange demonstrates an absolute requirement for Mg²⁺ and is optimal at an ADP/ATP ratio of 2. The high ATP concentration ($\text{K}{}_{0.5}\text{= 116 μ}\text{M}$) required for maximal exchange is interpreted as evidence for the involvement of a low affinity form of nucleotide site. The ATP/ADP exchange is regarded as evidence for an ADP-sensitive form of the phosphoenzyme. In native enzyme, pre-steady state kinetics show that the formation of the phosphoenzyme is partially sensitive to ADP while modification of the enzyme by pretreatment with 5,5′-dithiobis(2-nitrobenzoic acid) (DTNB) in the absence of Mg²⁺ results in a steady-state phosphoenzyme population, a component of which is ADP sensitive. The ATP/ADP exchange reaction can be either stimulated or inhibited by the presence of K⁺ as a function of pH and Mg²⁺.     

AutoFACT: AnAutomaticFunctionalAnnotation andClassificationTool

Background  

Assignment of function to new molecular sequence data is an essential step in genomics projects. The usual process involves similarity searches of a given sequence against one or more databases, an arduous process for large datasets.     

Characterization of cytochrome P-450SCC-containing liposomes

Purified cytochrome $\text{P}{}_{\mathrm{<mtext>450</mtext><mtext>SCC</mtext>}}$ from bovine adrenocortical mitochondria was incorporated into liposomes by the cholate-dilution method utilizing either dialysis or Sephadex gel filtration. Among synthetic phospholipids tested, dioleoylglycerophosphocholine showed the best stability during the incorporation of $\text{P}{}_{\mathrm{<mtext>450</mtext><mtext>SCC</mtext>}}$ into liposomes. A maximum amount of heme was incorporated into liposomes at a molar ratio of phospholipid to the cytochrome of approx. 200. When $\text{P}{}_{\mathrm{<mtext>450</mtext><mtext>SCC</mtext>}}$ was incorporated into the dioleoylglycerophosphocholine liposomes by the cholate-filtration method, the $\text{P}{}_{\mathrm{<mtext>450</mtext><mtext>SCC</mtext>}}\text{-}\text{containing liposomes}$ showed two major populations on the elution pattern of the Sepharose 4B gel filtration, and were seen at a diameter of 200–600 Å and its aggregated forms. When the cytochrome was incorporated into dioleoylglycerophosphocholine liposomes or cholesterol-free adrenocortical mitochondrial liposomes, $\text{P}{}_{\mathrm{<mtext>450</mtext><mtext>SCC</mtext>}}$ was less stable than $\text{P}{}_{\mathrm{<mtext>450</mtext><mtext>SCC</mtext>}}$ in aqueous solution. Cholesterol or adrenodoxin markedly stabilized the liposomal $\text{P}{}_{\mathrm{<mtext>450</mtext><mtext>SCC</mtext>}}$. Liposomal $\text{P}{}_{\mathrm{<mtext>450</mtext><mtext>SCC</mtext>}}$ required cholesterol for its optimum reduction with adrenodoxin, adrenodoxin reductase, and NADPH in the presence of CO. About 70% of the total heme in the dioleoylglycerophosphocholine liposomes was reduced by the enzymatic reduction in the presence of cholesterol, indicating that 70% of the total molecules are exposed to the surface of the outer monolayer. In order to see the location of the heme in membrane, the dioleoylglycerophosphocholine-liposomal $\text{P}{}_{\mathrm{<mtext>450</mtext><mtext>SCC</mtext>}}$ was subjected to $\text{p-}\text{chloromercuriphenyl sulfonic acid}$ treatment. This reagent destroyed the liposomal $\text{P}{}_{\mathrm{<mtext>450</mtext><mtext>SCC</mtext>}}$. These results suggest that the heme is located in the proximity of the $\text{p-}\text{chloromercuriphenyl sulfonic acid}$ reacting sites which are exposed to the surface, or located on the vincinity of polar heads of the membrane.     

Ligand-dependent reactivity of (Na+ + K+)-ATPase with showdomycin

Showdomycin inhibited pig brain ($\text{Na}{}^{\mathrm{+}}\text{+}\text{K}{}^{\mathrm{+}}\text{)-}\text{ATPase}$ with pseudo first-order kinetics. The rate of inhibition by showdomycin was examined in the presence of 16 combinations of four ligands, i.e., Na⁺, K⁺, Mg²⁺ and ATP, and was found to depend on the ligands added. Combinations of ligands were divided into five groups in terms of the magnitude of the rate constant; in the order of decreasing rate constants these were: (1)$\text{Na}{}^{\mathrm{+}}\text{+}\text{Mg}{}^{\mathrm{2+}}\text{+}\text{ATP}$, (2) $\text{Mg}{}^{\mathrm{2+}}$, $\text{Mg}{}^{\mathrm{2+}}\text{+}\text{K}{}^{\mathrm{+}}$, $\text{K}{}^{\mathrm{+}}$ and none, (3) $\text{Na}{}^{\mathrm{+}}\text{+}\text{Mg}{}^{\mathrm{2+}}\text{,}\text{Na}{}^{\mathrm{+}}$, $\text{K}{}^{\mathrm{+}}\text{+}\text{Na}{}^{\mathrm{+}}$ and $\text{Na}{}^{\mathrm{+}}\text{+}\text{K}{}^{\mathrm{+}}\text{+}\text{Mg}{}^{\mathrm{2+}}$, (4) $\text{Mg}{}^{\mathrm{2+}}\text{+}\text{K}{}^{\mathrm{+}}\text{+}\text{ATP}\text{,}\text{K}{}^{\mathrm{+}}\text{+}\text{ATP}$ and $\text{Mg}{}^{\mathrm{2+}}\text{+}\text{ATP}\text{, (5)}\text{K}{}^{\mathrm{+}}\text{+}\text{Na}{}^{\mathrm{+}}\text{+}\text{ATP}$, $\text{Na}{}^{\mathrm{+}}\text{+}\text{ATP}$, $\text{Na}{}^{\mathrm{+}}\text{+}\text{ATP}$, $\text{Na}{}^{\mathrm{+}}\text{+}\text{K}{}^{\mathrm{+}}\text{+}\text{Mg}{}^{\mathrm{2+}}\text{+}\text{ATP}$ and ATP. The highest rate was obtained in the presence of Na⁺, Mg²⁺ and ATP. The apparent concentrations of Na⁺, Mg²⁺ and ATP for half-maximum stimulation of inhibition ($\text{K}{}_{0.5}{}^{\mathrm{<mtext>s</mtext>}}$) were 3 mM, 0.13 mM and 4μM, respectively. The rate was unchanged upon further increase in Na⁺ concentration from 140 to 1000 mM. The rates of inhibition could be explained on the basis of the enzyme forms present, including E₁, E₂, ES, E₁-P and E₂-P, i.e., E₂ has higher reactivity with showdomycin than E₁, while E₂-P has almost the same reactivity as E₁-P. We conclude that the reaction of ($\text{Na}{}^{\mathrm{+}}\text{+}\text{K}{}^{\mathrm{+}}\text{)-}\text{ATPase}$ proceeds via at least four kinds of enzyme form (E₁, E₂, E₁ · nucleotide and EP), which all have different conformations.     

A/California/7/2009与A/California/4/2009病毒感染力比较

目的甲型H1N1流感病毒A/California/7/2009与A/California/4/2009病毒序列比较同源性在99%以上,本实验旨在比较两株病毒感染BALB/c小鼠研究感染力强弱。方法分别将A/California/7/2009（CA7）与A/California/4/2009（CA4）两株病毒分别连续10倍稀释后,对4~6周龄雌性BALB/c小鼠经乙醚麻醉后进行滴鼻攻毒,每个稀释度接种10只实验小鼠,测定CA7 MLD50为101.24/0.05 mL,检测小鼠感染、致病的多项指标,观察期为14 d。结果相同TCID50的CA7和CA4病毒感染小鼠,CA4感染小鼠后14 d内死亡率为20%,而CA7感染小鼠后8 d内死亡率为100%。CA7 106TCID50感染的小鼠病理表现为重度弥漫性间质性肺炎,CA4 106TCID50感染的小鼠病理表现为中度-重度间质性肺炎。结论在相同条件下,CA7感染力明显强于CA4。     

Characterization of partially purified (Na+ + K+)-ATPase from porcine lens

The partial purification of $\text{(}\text{Na}{}^{\mathrm{+}}\text{+}\text{K}{}^{\mathrm{+}}\text{)-}\text{ATPase}$ from pig lens has been achieved by treatment with deoxycholate followed by density gradient centrifugation. The specific activity of the final preparation, ranging from 300 to 500 nmol/h per mg protein, is increased approx. 100-fold compared to the homogenate. A parallel increase in $\text{p-}\text{nitrophenylphosphatase}$ activity is also observed. Sodium dodecyl sulfate (SDS) gel electrophoresis reveals six major protein bands, one of which is the 93 kDa α subunit of $\text{(}\text{Na}{}^{\mathrm{+}}\text{+}\text{K}{}^{\mathrm{+}}\text{)-}\text{ATPase}$ which can be phosphorylated by reaction with [γ-³²P]ATP. A second band contains a glycoprotein which displays an apparent molecular weight of 51 000 and thus appears to be the β subunit of the enzyme. The enzyme is sensitive to ouabain with the $\text{I}{}_{50}$ for $\text{(}\text{Na}{}^{\mathrm{+}}\text{+}\text{K}{}^{\mathrm{+}}\text{)-}\text{ATPase}$ and $\text{p-}\text{nitrophenylphosphatase}$ inhibition being 1.2 and 1.3 μM, respectively. Several agents which inhibit $\text{Na}{}^{\mathrm{+}}\text{+}\text{K}{}^{\mathrm{+}}\text{)-}\text{ATPase}$ from other tissues such as oligomycin, Ca²⁺, vanadate, $\text{N-}\text{ethylmaleimide}$, $\text{p-}\text{chloromercuribenzenesulfonic acid}$ (PCMBS) and 5,5′-dithiobis-(2-nitrobenzoic acid) (DTNB) also inhibit the lens enzyme. Monovalent cations other than K⁺ are partially effective in activating the ($\text{Na}{}^{\mathrm{+}}\text{+}\text{K}{}^{\mathrm{+}}$)-ATPase and $\text{p-}\text{nitrophenylphosphatase}$ activities. The K⁺ congeners were relatively more effective in supporting $\text{(}\text{Na}{}^{\mathrm{+}}\text{+}\text{K}{}^{\mathrm{+}}\text{)-}\text{ATPase}$ compared to $\text{p-}\text{nitrophenylphosphatase}$ activity. Other kinetic properties of the lens enzyme are also comparable to those of the enzyme from other tissues. Utilizing the partially purified membrane bound enzyme, discontinuities in Arrhenius plots of $\text{(}\text{Na}{}^{\mathrm{+}}\text{+}\text{K}{}^{\mathrm{+}}\text{)-}\text{ATPase}$ activity, $\text{p-}\text{nitrophenylphosphatase}$ activity and fluoresence polarization of the fluidity probe, 1,6-diphenyl-1,3,5-hexatriene (DPH), are observed near the physiological temperature of lens. The possible significance of these observations for the mechanism of cataract formation are discussed.     

An electron spin probe study of (Na+ + K+)-ATPase-Containing membranes

The modulating effect of membrane lipids on enzyme function has been described by several investigators. We have used the spin probe $\text{N-}\text{oxyl}\text{-4′,4′-}\text{dimethyloxazolidine}\text{-12-}\text{keto}$ methyl stearate (M 12-NSE) to study this interaction in ox brain membranes enriched with $\text{(}\text{Na}{}^{\mathrm{+}}\text{+}\text{K}{}^{\mathrm{+}}\text{)-}\text{ATPase}$. This methyl ester of stearic acid is practically insoluble in aqueous media, and consequently spectra of M 12-NSE-labelled preparations are free of “liquid lines”.At least two types of spectra may be obtained when ox brain microsomes are spin labelled with M 12-NSE, indicating the presence of two distinct binding sites. At one site the spin label is relatively unrestricted and gives rise to an isotropic spectrum. A second spectrum, which is obtained from spin label at another site, is similar to that which is observed after incorporation of M 12-NSE into phospholipid bilayers. This suggests that this latter site is within the core of the microsomal membrane.The two binding sites differ in their affinity for the spin probe. The low affinity site is both more abundant in crude preparations and is more easily removed by detergent treatment; spin labels at this site produce isotropic spectra. The high affinity sites are fewer in number and produce broad spectra. In addition these high affinity sites increase in concentration as the enzyme undergoes purification.The two sites are quite distinct in their sensitivity to ascorbic acid, the low affinity site showing a considerably greater rate of reduction by this agent.This study also demonstrates that the delipidation effects of sodium dodecyl sulfate and sodium deoxycholate on $\text{(}\text{Na}{}^{\mathrm{+}}\text{+}\text{K}{}^{\mathrm{+}}\text{)-}\text{ATPase-enriched}$ microsomes from ox brain are not identical.It is suggested that the two spin probe binding sites represent two different lipid domains, one of which is very closely associated with the $\text{(}\text{Na}{}^{\mathrm{+}}\text{+}\text{K}{}^{\mathrm{+}}\text{)-}\text{ATPase}$ enzyme and may reflect a protein-directed phospholipid specificity for this enzyme.     

Annular and non-annular binding sites on the (Ca2+ + Mg2+)-ATPase

Quenching of the fluorescence of the $\text{(}\text{Ca}{}^{\mathrm{2+}}\text{+}\text{Mg}{}^{\mathrm{2+}}\text{)-}\text{ATPase}$ purified from muscle sarcoplasmic reticulum can be used to measure relative binding constants of hydrophobic compounds to the phospholipid-protein interface. We show that the binding constant for cholesterol is considerably less than that for phosphatidylcholine, so that cholesterol is effectively excluded from the phospholipid annulus around the ATPase. However, dibromocholestan-3β-ol causes quenching of the fluorescence of the ATPase, and so has access to other, non-annular sites. We suggest that these non-annular sites could be at protein/protein interfaces in ATPase oligomers. Oleic acid can bind at the phospholipid/protein interface, although its binding constant is less than that for a phosphatidylcholine, and it can also bind at the postulated non-annular sites. The effects of these compounds on the activity of the ATPase depend on the structure of the phospholipid present in the systems.     

Comparison of parameters estimated from A/Ci and A/Cc curve analysis

The parameters estimated from traditional A/C _i curve analysis are dependent upon some underlying assumptions that substomatal CO₂ concentration (C _i) equals the chloroplast CO₂ concentration (C _c) and the C _i value at which the A/C _i curve switches between Rubisco- and electron transport-limited portions of the curve (C _i-t) is set to a constant. However, the assumptions reduced the accuracy of parameter estimation significantly without taking the influence of C _i-t value and mesophyll conductance (g _m) on parameters into account. Based on the analysis of Larix gmelinii’s A/C _i curves, it showed the C _i-t value varied significantly, ranging from 24 Pa to 72 Pa and averaging 38 Pa. t-test demonstrated there were significant differences in parameters respectively estimated from A/C _i and A/C _c curve analysis (p<0.01). Compared with the maximum ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) carboxylation rate (V_cmax), the maximum electron transport rate (J_max) and J_max/V_cmax estimated from A/C _c curve analysis which considers the effects of g _m limit and simultaneously fits parameters with the whole A/C _c curve, mean V_cmax estimated from A/C _i curve analysis (V_cmax-C _i) was underestimated by 37.49%; mean Jmax estimated from A/C _i curve analysis (J_max-C _i) was overestimated by 17.8% and (J_max-C _i)/(V_cmax-C _i) was overestimated by 24.2%. However, there was a significant linear relationship between V_cmax estimated from A/C _i curve analysis and V_cmax estimated from A/C _c curve analysis, so was it J_max (p<0.05).     

Regulation of rat brain (Na+ + K+)-ATPase activity by cyclic AMP

The interaction between the $\text{(}\text{Na}{}^{\mathrm{+}}\text{+}\text{K}{}^{\mathrm{+}}\text{)-}\text{ATPase}$ and the adenylate cyclase enzyme systems was examined. Cyclic AMP, but not 5′-AMP, cyclic GMP or 5′-GMP, could inhibit the $\text{(}\text{Na}{}^{\mathrm{+}}\text{+}\text{K}{}^{\mathrm{+}}\text{)-}\text{ATPase}$ enzyme present in crude rat brain plasma membranes. On the other hand, the cyclic AMP inhibition could not be observed with purified preparations of $\text{(}\text{Na}{}^{\mathrm{+}}\text{+}\text{K}{}^{\mathrm{+}}\text{)-}\text{ATPase}$ enzyme. Rat brain synaptosomal membranes were prepared and treated with either NaCl or cyclic AMP plus NaCl as described by Corbin, J., Sugden, P., Lincoln, T. and Keely, S. ((1977) J. Biol. Chem. 252, 3854–3861). This resulted in the dissociation and removal of the catalytic subunit of a membrane-bound cyclic AMP-dependent protein kinase. The decrease in cyclic AMP-dependent protein kinase activity was accompanied by an increase in $\text{(}\text{Na}{}^{\mathrm{+}}\text{+}\text{K}{}^{\mathrm{+}}\text{)-}\text{ATPase}$ activity. Exposure of synaptosomal membranes containing the cyclic AMP-dependent protein kinase holoenzyme to a specific cyclic AMP-dependent protein kinase inhibitor resulted in an increase in $\text{(}\text{Na}{}^{\mathrm{+}}\text{+}\text{K}{}^{\mathrm{+}}\text{)-}\text{ATPase}$ enzyme activity. Synaptosomal membranes lacking the catalytic subunit of the cyclic-AMP-dependent protein kinase did not show this effect. Reconstitution of the solubilized membrane-bound cyclic AMP-dependent protein kinase, in the presence of a neuronal membrane substrate protein for the activated protein kinase, with a purified preparation of $\text{(}\text{Na}{}^{\mathrm{+}}\text{+}\text{K}{}^{\mathrm{+}}\text{)-}\text{ATPase}$, resulted in a decrease in overall $\text{(}\text{Na}{}^{\mathrm{+}}\text{+}\text{K}{}^{\mathrm{+}}\text{)-}\text{ATPase}$ activity in the presence of cyclic AMP. Reconstitution of the protein kinase alone or the substrate protein alone, with the $\text{(}\text{Na}{}^{\mathrm{+}}\text{+}\text{K}{}^{\mathrm{+}}\text{)-}\text{ATPase}$ has no effect on $\text{(}\text{Na}{}^{\mathrm{+}}\text{+}\text{K}{}^{\mathrm{+}}\text{)-}\text{ATPase}$ activity in the absence or presence of cyclic AMP. Preliminary experiments indicate that, when the activated protein kinase and the substrate protein were reconstituted with the $\text{(}\text{Na}{}^{\mathrm{+}}\text{+}\text{K}{}^{\mathrm{+}}\text{)-}\text{ATPase}$ enzyme, there appeared to be a decrease in the Na⁺-dependent phosphorylation of the Na⁺-ATPase enzyme, while the K⁺-dependent dephosphorylation of the $\text{(}\text{Na}{}^{\mathrm{+}}\text{+}\text{K}{}^{\mathrm{+}}\text{)-}\text{ATPase}$ was unaffected.