复杂网络分析激酶底物信号传递机制

为了研究信号从激酶到达底物蛋白质后的传递机制,本文将磷酸化和蛋白质相互作用结合构建网络,利用复杂网络理论对该网络进行分析.结果发现,底物中存在的社区结构不会只传递特定激酶组的信号,影响网络稳定性的关键节点大多参与肿瘤发生相关的信号途径.     

Cardiac protective role of a novel erythrocyte-derived depressing factor on rats and its Ca~(2+) mechanism

Erythrocyte-derived depressing factor (EDDF) has been extracted from human erythrocytes by our lab for the first time. Our previous research has shown that EDDF is different from those known blood pressure modulators in the body. It may be a new endogenous player. The physiological functions, toxicity, physical and biological properties, existence and tissue distribution as well as biochemical structure have been well studied in our lab[1—4]. Our previous findings indicated that EDDF p…     

微波法制备白芨多糖磷酸酯

以磷酸二氢钠和磷酸氢二钠混合磷酸盐为磷酰化试剂,尿素为催化剂,微波辅助制备白芨多糖磷酸酯,试图寻找一种方便有效的多糖磷酸酯制备方法.考察了反应温度、微波辐射时间、磷酸盐总量和溶液pH值对反应的影响.得到最佳反应条件为:反应温度50°C,pH值为5,辐射时间3min,磷酸盐用量50%,所得产物中磷取代度为1.57%.微波制备方法的反应温度低、时间短、反应物用量小、产物的取代度高,是一种高效的白芨多糖磷酸酯合成方法.     

使用多样性增量预测磷酸化位点

磷酸化是蛋白质最重要的翻译后修饰之一.应用基于多样性增量的二次判别分析 (Increment of Diversity with Quadratic Discriminant analysis,IDQD)方法对CK2,PKA和PKC三种类型磷酸化位点进行预测,k-fold交叉检验的正确率分别为86%,90%和85%,独立测试集检验的正确率分别为86%,88%和84%.所得结果高于包括支持向量机在内的现有预测方法.     

磷酸化处理对蚕蛹多肽结合钙能力的影响

为探索磷酸化处理对多肽结合钙能力的促进作用,以蚕蛹多肽为研究对象,采用不同种类和添加量的磷酸化试剂对其进行磷酸化改性,再与CaCl₂反应制备蚕蛹多肽结合钙复合物,并对样品进行钙和磷含量测定、磷酸化位点检测、红外光谱分析和扫描电镜分析。研究结果表明:Na₃PO₄、NaH₂PO4、(NaPO₃)₆ 3种试剂可与蚕蛹多肽发生磷酸化反应,改性后蚕蛹多肽结合钙含量可从145.12mg/g提高至202.32mg/g ;不同种类磷酸化试剂处理对多肽的结合钙能力无显著性差异,但磷酸化程度对肽结合钙能力有较大影响;多肽结合钙复合物中钙含量随磷酸化试剂的增加先升高后下降,当采用质量分数为0.50%的NaH₂PO₄时,改性后蚕蛹多肽结合钙含量达到最大值。红外光谱和磷酸化位点检测表明:NaH₂PO₄处理可使蚕蛹酶解产物中的17种肽段被磷酸化,增加磷酸化位点22个;肽段主要通过与丝氨酸和苏氨酸上的—OH反应接入磷酸基团。研究表明,磷酸化改性方法可显著提高多肽结合钙能力,希望为高效的补钙制品开发提供新的思路和方法。     

Phosphorylation of cockroach antennal polypeptides: effects of second messengers and pheromonal blend

In insect antennal extracts, Schleicher et al.¹ showed that protein kinase C (PKC) inhibitors abolish the transience of pheromone-induced rapid inositol trisphosphate responses, which suggests that pheromonal signals act on phosphorylation of specific proteins. To confirm this hypothesis, we studied the effects of second messengers and a pheromonal blend on phosphorylation of antennal proteins in the cockroachPeriplaneta americana. Proteins from adult male antennae were phosphorylated in vitro in the presence of [³²P] triphosphate, then separated by SDS-polyacrylamide gel electrophoresis. Numerous phosphopolypeptides were visualized. The presence of Ca⁺⁺/calmodulin in the incubation medium resulted in increased phosphorylation of polypeptides with molecular weights of 38, 48, 51, 54 and 58 kDa. Stimulation of PKC by addition of Ca⁺⁺ phosphatidylserine (PS)/phorbol myristate acetate (PMA) resulted in the appearance of three phosphopolypeptides of 36, 70 and 120 kDa. In the presence of cyclic adenosine monophosphate, two new major polypeptides of 46 and 42 kDa appeared; the latter polypeptide also appeared in the presence of cyclic guanosine monophosphate. Comparison with polypeptide composition of tissue from the cerci, leg, brain and fat body showed that the 36 and 48 kDa polypeptides were specific to antennae, whereas the 120 kDa polypeptide was also present in the adult brain. When antennae are subjected to pheromonal stimulation for 16 seconds prior to homogenization, in vitro phosphorylation of the 120, 70, 64 and 38 kDa polypeptides was inhibited, whereas phosphorylation of the 58, 54, 51 and 48 kDa polypeptides was strongly stimulated. It is noteworthy that a 107 kDa polypeptide was observed only after pheromonal stimulation by Ca⁺⁺/PS/PMA. Our findings suggest that Ca⁺⁺-and PKC-dependent protein phosphorylation systems play an important role in the transduction of pheromonal signals in antennae of male cockroachP. americana. We speculate that specific phosphoproteins may modulate sensitivity and signal amplification during the olfactory transduction process.     

组胺能与γ-氨基丁酸能神经传递在间位核神经元信息加工中的作用(英文)

小脑间位核(interpositus nucleus,IN)主要接受γ-氨基丁酸(GABA)能纤维支配,同时接受组胺能纤维的调节.本研究在小脑脑片上研究了GABA和组胺对单个IN神经元电活动的共同作用.持续灌流组胺或同时施加组胺和GABA,81.2%(69/85)神经元,GABA及其激动剂的效应都被组胺削弱(持续灌流n=33;同时施加n=36).这种削弱效应能够被组胺H2受体阻断剂ranitidine(n=10)和PKA抑制剂H-89阻断(n=8),fors-kolin模拟组胺的效应(n=9).结果表明组胺和GABA对IN神经元的电活动具有交互调节作用:通过激活H2受体偶联的G-protein-AC-PKA信号通路,磷酸化GABAB和GABAA受体,降低受体功能.推测受体间的对话的工作模式,可能是整个大脑神经元活动的某些药理作用和生理活动调节的基础;如果对话紊乱,可能导致大脑功能障碍.     

Research progress in protein post-translational modification

A. Ciechanover and A. Hershko, Israel scientists, and American scientist O. Rose have received the Nobel Prize of 2004 in chemistry because they have discovered themechanism of the ubiquitin-regulated proteolysis. Their work indicates the research directi…     

蛋白质的磷酸化修饰及其研究方法

蛋白质磷酸化是一种重要的翻译后修饰，它参与和调控生物体内的许多生命活动。随着蛋白质组技术的不断发展，蛋白质磷酸化的研究越来越受到广泛的重视。本文介绍了蛋白质磷酸化修饰的主要类型与功能、磷酸化兰白质及磷酸化肽的标记和分离与富集、磷酸化肽及磷酸化位点分析以及蛋白质的磷酸化改性等方法,并综述了近年来国内外的主要研究进展。     

The ATP synthase (F0−F1) complex in oxidative phosphorylation

The transmembrane electrochemical proton gradient generated by the redox systems of the respiratory chain in mitochondria and aerobic bacteria is utilized by proton translocating ATP synthases to catalyze the synthesis of ATP from ADP and P_i. The bacterial and mitochondrial H⁺-ATP synthases both consist of a membranous sector, F₀, which forms a H⁺-channel, and an extramembranous sector, F₁, which is responsible for catalysis. When detached from the membrane, the purified F₁ sector functions mainly as an ATPase. In chloroplasts, the synthesis of ATP is also driven by a proton motive force, and the enzyme complex responsible for this synthesis is similar to the mitochondrial and bacterial ATP synthases. The synthesis of ATP by H⁺-ATP synthases proceeds without the formation of a phosphorylated enzyme intermediate, and involves co-operative interactions between the catalytic subunits.