丝瓜种子萌芽及子叶生长过程中谷氨酰胺合成酶同工酶的变化

测定了丝瓜种子萌芽及子叶不同生长阶段的谷氨酰胺合成酶(GS)的活性及其同工酶的变化．除了发育上的变化外，光和外源氮对GS活性及其同工酶影响也是显著的．无论外源氮(N)是否存在，GS活性在种子萌芽和子叶生长初期是逐渐升高的，于第6天达到最大，其后活性逐步降低．在光照下，外源氮对GS1诱导作用大于GS2；在无氮下，光对GS2的刺激大于GS1；在暗处，外源氮对GS1和GS2两者都有诱导作用．     

Cloning and Expression of Rat Liver S-Adenosylmethionine Synthetase

Introduction S Adenosylmethionine(SAM)isabiologicallyac tivecompoundwidelydistributedinthebodytissues andfluids.Itisinvolvedinanumberofbiochemical reactions.Asamethylgroupdonor,itisimportantin transmethylationreactions,whichcontributestothe synthesesofsuc…     

Simultaneous measurement of glucose and glutamine in aqueous solutions by near infrared spectroscopy

A method is described for measuring the concentrations of both glucose and glutamine in binary mixtures from near infrared (NIR) absorption spectra. Spectra are collected over the range from 5000–4000/cm (2.0–2.5μm) with a 1-mm optical path length. Glucose absorbance features at 4710, 4400, and 4300/cm and glutamine features at 4700, 4580, and 4390/cm provide the analytical information required for the measurement. Multivariate calibration models are generated by using partial least squares (PLS) regression alone and PLS regression combined with a preprocessing digital Fourier filtering step. The ideal number of PLS factors and spectral range are identified separately for each analyte. In addition, the optimum Fourier filter parameters are established for both compounds. The best overall analytical performance is obtained by combining Fourier filtering and PLS regression. Glucose measurements are established over the concentration range from 1.66–59.91 mM, with a standard error of prediction (SEP) of 0.32 mM and a mean percent error of 1.84%. Glutamine can be measured over the concentration range from 1.10–30.65 mM with a SEP of 0.75 mM and a mean percent error of 6.67%. These results demonstrate the analytical utility of NIR spectroscopy for monitoring glucose and glutamine levels in mammalian and insect cell cultures.     

谷氨酸钠从其水溶液中的周期结晶现象

早在本世纪初物理化学家们就已观察到，某些物质从其熔融态或溶液中结晶出来时，在某些条件下可形成周期性的宏观结晶条纹［1，2］，这就是周期结晶现象，由于这种现象和晶体生长、材料物性以及成矿过程等重要问题紧密相关，很早就引起了人们的重视．近年来有关非平衡非线性系统中自发产生各种时空有序现象（自组织现象）的各种理论（如耗散结构理论）［3］的兴起，重新引起了人们对这类现象的兴趣．八十年代，Iwamoto等先后报道了甲基苄基氨基甲酸酯（MethylMesitylcarbamate）从氯仿溶液［4］和抗坏血酸（AscorbicAcid）从甲醇溶液［5］…     

ALLOSTERIC REGULATION OF THE STATE OF ADENYLYLATION OF GLUTAMINE SYNTHETASE IN PERMEABILIZED CELL PREPARATIONS OF Rhoaopseudomonas sphaeroides

Following a freeze-thaw cycle, and the treatment of Rhodopseudomonas sphaeroides withthe nonionic detergent Lubrol PX, the permeabilized cell suspensions can be assayed directlyboth for the intracellular levels of glutamine synthetase and the state of adenylylation(i.e. the average number n of adenylylated subunits/dodecameric molecules). It seems that all components of the bicycle system are retained if cells grown with lowconcentrations of a?monia as the sole nitrogen source are used. The value of n was depend-ent upon the concentration of substrates (ATP, Pi) and allosteric effectors (ATP, gluta-mine and α-ketoglutarate) of adenylyltransferase. The value of n affected by UTP, thespecific substrate of the uridylyltransferase shows first the evidence that the bicycle cascadecontrol system studied in Escherichia coli may exist in this phototrophic bacterium.     

丙氨酰-谷氨酰胺的简便有效合成

谷氨酰胺 (Ｇｌｎ)是肠道必需氨基酸 ,是肠粘膜细胞氧化的重要燃料。肠粘膜缺乏Ｇｌｎ将严重损伤肠屏障功能 ,创伤性休克及化疗应激可加重损害 ,导致肠粘膜萎缩 ,肠道细菌移位 ,甚至促发脓毒症和多器官功能不全 (ＭＯＤＳ) [1] 。但Ｇｌｎ性质不稳定 ,不能耐高温消毒 ,因此 ,一般氨基酸制剂不含Ｇｌｎ。Ａｌａ Ｇｌｎ为Ｇｌｎ代谢前体 ,溶解度高 ,性质较Ｇｌｎ稳定 ,能耐受热灭菌 ,在体内数分钟即分解出Ｇｌｎ ,能部分补偿肠道所需Ｇｌｎ ,减少严重创伤及腹腔注射化疗药物 5 氟尿嘧啶 (5 ＦＵ)引发的小肠粘膜形态功能改变 ,细菌移位 ,降低…     

Growth inhibition in animal cell culture

Eight independent cell lines accumulated ammonia in culture to concentrations between 1.3 and 2.9 mM. The growth inhibition of such concentrations of ammonium chloride when added to culture medium was variable. The cell lines tested could be divided into 3 groups depending on their growth response to 2 mM added NH4Cl. In the first group (293, HDF, Vero, and PQXB1/2) little (less than 14%) or no growth inhibition occurred. In the second group (McCoy and MDCK) a reduction in final cell yield of 50-60% was observed. The third group (HeLa and BHK) was most sensitive to the effects of NH4Cl with growth inhibition (greater than 75%) compared to controls. The growth inhibitory effect of added lactate up to 20 mM was negligible (less than 10%) for 3 cell lines, although one cell line (PQXB1/2) showed greater sensitivity. The interactive effects of ammonia and lactate were determined in a matrix experiment. At lactate (greater than 12 mM) and ammonia (1-4 mM), the growth inhibitory effects of the two components were synergistic. However, at low concentrations of lactate (less than 12 mM) the toxic effect of ammonia was reduced. A proposed mechanism for the sparing effect of lactate on ammonia toxicity is discussed. This may have importance in developing strategies for the optimal growth of ammonia-sensitive cell lines.     

Chemoenzymatic synthesis of β-carboline derivatives using McbA,a new ATP-dependent amide synthetase

McbA was characterized in vitro as a novel amide synthetase in the marinacarbolines A–D biosynthetic pathway, catalyzing amide bond formation between 1-acetyl-3-carboxy-β-carboline (1a) and substituted-β-phenethylamines (1b, 2b, 3b) and tryptamine (4b) in an ATP-dependent manner. Enzyme kinetic analyses highlight β-phenethylamine as the most suitable amine donor. McbA showed broad substrate compatibility with substituted amines; 10 new β-carboline analogues were chemoenzymatically generated.     

Metabolic Profiling of Amino Acids by Liquid Chromatography–Tandem Mass Spectrometry (LC–MS) to Characterize the Significance of Glutamine in Triple-Negative Breast Cancer (TNBC)

Triple-negative breast cancer (TNBC) is considered to be aggressive based on its low overall survival and disease-free rates. Currently, there is no molecular-targeted therapy. The identification of a suitable biomarker is still a research focus for TNBC at the present time. Amino acid metabolism fulfills multiple important physiological roles in humans. Their metabolic abnormalities have been reported in numerous cancer studies and amino acid pathways may also be chemotherapeutic targets. This study reports the profiling analysis of amino acids in TNBC and non-TNBC cell lines for detecting biomarkers based on the strategy of N-phosphorylation labeling with liquid chromatography–tandem mass spectrometry (LC–MS). Glutamine (Gln) was found to be significantly down-regulated in TNBC cells because it was largely absorbed and consumed in the TNBC cell lines. These results indicate faster proliferation of TNBC and higher consumption of glutamine to meet the material and energy demand, suggesting its potential role in TNBC progression. Hence, glutamine may be regarded as a biomarker and Gln-targeted approaches may become a new therapeutic strategies for TNBC.     

4-氨基-N8,N10-二碳杂-N5-取代四氢叶酸类似物的合成及生物活性研究

以2,4-二氨基-6-羟甲基吡啶并[3,2-d]嘧啶为原料,与对甲酰基苯甲酰谷氨酸二乙酯发生Wittig反应构建叶酸类似物的骨架结构,还原吡啶环得到4-氨基-8,10-二碳杂四氢叶酸类似物,在N5位取代不同基团得到四个新的4-氨基-N8,N10-二碳杂四氢叶酸类化合物,经1H NMR,13C NMR和MS对化合物的结构进行了表征.初步生物活性结果表明,此类化合物对人重组胸苷酸合成酶的抑制作用与N5位取代基有关,2个化合物在0.1 μmol·L-1的浓度下对HL-60白血病细胞的抑制率达到60％以上.