水杨酸处理诱导采后甜瓜抗坏血酸-还原型谷胱甘肽循环代谢清除过氧化氢的作用及机制

目的：研究抗坏血酸（ascorbic acid，AsA）-还原型谷胱甘肽（reduced glutathione，GSH）循环代谢在水杨酸处理采后甜瓜诱导的过量H2O2清除过程中的作用。方法：用4 mmol/L水杨酸浸泡‘玉金香’厚皮甜瓜10 min，测定处理后果实常温贮藏过程中丙二醛（malondialdehyde，MDA）含量，分析活性氧的积累水平、超氧化物歧化酶（superoxide dismutase，SOD）和过氧化氢酶（catalase，CAT）活力，以及AsA-GSH循环过程相关酶活力及产物和底物含量。结果：水杨酸处理降低了果实MDA含量，第10天处理组MDA含量较对照组降低14.6%；显著提高了果实O2－·的产生速率和H2O2含量（P＜0.05），其中处理后第2天O2－·的产生速率高出同期对照组的1.9 倍，第6天H2O2含量高出对照组果实29.7%；提高了果实SOD活力，但抑制了CAT活力，说明H2O2的清除可能是依赖于除酶促系统外的其他系统。此外，水杨酸处理提高了果实抗坏血酸过氧化物酶、单脱氢抗坏血酸过氧化物酶、脱氢抗坏血酸还原酶和谷胱甘肽还原酶的活力，增加了AsA和氧化型谷胱甘肽的含量，降低了脱氢抗坏血酸和GSH的含量。结论：水杨酸处理诱导了厚皮甜瓜果实的氧爆，抑制了MDA产生，由水杨酸诱导产生的过量H2O2主要依靠AsA-GSH循环系统清除。     

基于氮掺杂碳纳米颗粒荧光猝灭-恢复方法检测还原型谷胱甘肽

分别以海藻酸钠和色氨酸为碳源和氮源,采用固相法一步合成出了量子产率为47.9%的氮掺杂荧光碳纳米颗粒(N-CNPs)。根据铜离子存在的条件下,N-CNPs荧光强度的恢复情况与还原型谷胱甘肽浓度成正比的关系,建立基于N-CNPs检测还原型谷胱甘肽的新方法。优化了溶液pH及反应时间等条件。在pH 6.0、铜离子浓度30μmol/L条件下,谷胱甘肽在0.2~45μmol/L浓度范围内与N-CNPs荧光恢复强度呈良好的线性关系,检出限为50 nmol/L。方法具有灵敏度高、选择性好、操作简单等优点,可用于实际样品中谷胱甘肽的检测。     

Induction of favism-like symptoms in the rat: Effects of vicine and divicine in normal and buthionine sulphoximine-treated rats

Rats were fed an adequate or a deficient diet and offered water or buthionine sulphoximine (BSO) solution for 2 weeks, and then the same diets with vicine for another week in experiment 1. BSO in combination with the deficient diet caused a marked decrease in blood glutathione (GSH) and growth retardation but failed to show any effects resulting from supplementation with vicine. In experiment 2 the rats were given an adequate diet and BSO as before, and injected intravenously with divicine (DV). Here again, BSO depressed rat growth, and so did DV. Each of the insults also caused haematological changes, especially a fall in GSH, but the most severe changes appeared in the group treated with both BSO and DV. A decrease in haematocrit and increases in adrenal and spleen weight were also noted. In experiment 3 the rats were injected with different doses of DV, without pretreatment with BSO. The main effect was a drop in blood GSH and haematocrit, and an increase in adrenal and spleen weights, all of which were dose-related. Administration of the higher doses of DV resulted in a severe cyanosis followed by death within a relatively short period of time.     

Multimer formation as a consequence of separate homodimerization domains: the human c-Jun leucine zipper is a transplantable dimerization module

Human c-Jun and c-Fos leucine zipper domains were examined fortheir ability to serve as autonomous dimerization domains aspart of a heterologous protein construct. Schistosoma japonicumglutathione S-transferase (GST) was fused to recombinant Junleucine zipper (rJunLZ) and Fos leucine zipper (rFosLZ) domains.SDS–PAGE ‘snapshot’ analyses based on disulphidelinkage of monomers demonstrated the ability of rJunLZ to functionas a dimerization motif in a foreign protein environment. Sterichindrance prevented formation of rJunLZ–GST::rFosLZ–GSTheterodimers whereas rJunLZ–GST::rFosLZ and rJunLZ::rFosLZ–GSTformed readily. Furthermore, rJunLZ–GST generated homodimerssuggesting fusion protein heterodimers interact differentlyto homodimers. Gel filtration chromatography confirmed thatGST is a dimer in solution and that attachment of a leucinezipper domain allows further interactions to take place. Sedimentationequilibrium analyses showed that GST is a stable dimer (K_a >10⁶ M^-1) with no higher multimeric forms. rFosLZ–GST weaklyassociates beyond a dimer (K_a {small tilde}4x10⁵ M^-1) and rJunLZ–GSTassociates indefinitely (K_a {small tilde}4x10⁶ M^-1), consistentwith an isodesmic model of association. The interaction of theseleucine zippers independently of GST association demonstratestheir utility in the modification of proteins when multimerformation is desired.     

多肽配基亲和吸附介质的配基密度控制规律

以谷胱甘肽为配基,琼脂糖微球为骨架,探索将谷胱甘肽通过共价键偶联到琼脂糖微球骨架上,制备可以分离谷胱甘肽S转移酶(Glutathione S-transferase, GST)及以其为标签的融合蛋白的亲和吸附介质.采用正交实验方法考察了谷胱甘肽加入量、偶联缓冲液的pH值和反应温度对亲和介质配基密度的影响.结果发现,该反应过程中的pH值对配基密度影响最大,其次为谷胱甘肽的加入量.用所制备的亲和吸附介质纯化GST(大鼠肝脏谷胱甘肽S转移酶),发现GST的吸附量随配基密度增加而增加,但GST活性却随配基密度的增加而下降,较好的干胶配基密度为260 μmol/g.大鼠肝匀浆液经过离子交换和亲和层析两个步骤,获得了电泳纯的GST,比活力为12.08 U/mg,总活性回收率为40%以上.     

邻苯二甲醛同步荧光衍生化法测定微量指血中还原型谷胱甘肽

采用微升（μL）级微量指血为检测样品,在未对样品去蛋白的条件下,用邻苯二甲醛（OPA）荧光衍生化法测定了微量静脉血中还原型谷胱甘肽（GSH）的含量.利用标准曲线法和标准加入法进行3次测定的均值分别为4.415μmol/L和5.417 5μmol/L,加标回收率为95.28%～97.18%,检测限为3.6×10-8 mol/L,可基本满足微量成分分析所要求的准确度.本研究为灵敏、简便和快速测定微量血样中GSH的含量提供了实验和方法学的依据.     

还原型谷胱甘肽提取方法初探

还原型谷胱甘肽(GSH)是一种天然的活性短肽，具有清理体内自由基等生理功能。为了寻找适合实验室应用的简便、易行的提取GSH的方法，以安琪活性干酵母为研究对象，以相似相容原理为理论依据，用热水抽提、甲酸抽提、乙醇抽提、低温抽提方法提取GSH，分析比较不同提取方法的提取效果。结果表明，热水抽提是一种比较理想的从酵母中提取谷胱甘肽的方法，其特点是耗时少、仪器简单、抽提液中GSH质量浓度高、经济且无污染。     

Water-gated phthalocyanine transistors: Operation and transduction of the peptide–enzyme interaction

The use of aqueous solutions as the gate medium is an attractive strategy to obtain high charge carrier density (10¹² cm⁻²) and low operational voltages (<1 V) in organic transistors. Additionally, it provides a simple and favorable architecture to couple both ionic and electronic domains in a single device, which is crucial for the development of novel technologies in bioelectronics. Here, we demonstrate the operation of transistors containing copper phthalocyanine (CuPc) thin-films gated with water and discuss the charge dynamics at the CuPc/water interface. Without the need for complex multilayer patterning, or the use of surface treatments, water-gated CuPc transistors exhibited low threshold (100 ± 20 mV) and working voltages (<1 V) compared to conventional CuPc transistors, along with similar charge carrier mobilities (1.2 ± 0.2) x 10⁻³ cm² V⁻¹ s⁻¹. Several device characteristics such as moderate switching speeds and hysteresis, associated with high capacitances at low frequencies upon bias application (3.4–12 μF cm⁻²), indicate the occurrence of interfacial ion doping. Finally, water-gated CuPc OTFTs were employed in the transduction of the biospecific interaction between tripeptide reduced glutathione (GSH) and glutathione S-transferase (GST) enzyme, taking advantage of the device sensitivity and multiparametricity.