Metabolism of Pentose Sugars in the Hyperthermophilic Archaea Sulfolobus solfataricus and Sulfolobus acidocaldarius

We have previously shown that the hyperthermophilic archaeon, Sulfolobus solfataricus, catabolizes d-glucose and d-galactose to pyruvate and glyceraldehyde via a non-phosphorylative version of the Entner-Doudoroff pathway. At each step, one enzyme is active with both C6 epimers, leading to a metabolically promiscuous pathway. On further investigation, the catalytic promiscuity of the first enzyme in this pathway, glucose dehydrogenase, has been shown to extend to the C5 sugars, d-xylose and l-arabinose. In the current paper we establish that this promiscuity for C6 and C5 metabolites is also exhibited by the third enzyme in the pathway, 2-keto-3-deoxygluconate aldolase, but that the second step requires a specific C5-dehydratase, the gluconate dehydratase being active only with C6 metabolites. The products of this pathway for the catabolism of d-xylose and l-arabinose are pyruvate and glycolaldehyde, pyruvate entering the citric acid cycle after oxidative decarboxylation to acetyl-coenzyme A. We have identified and characterized the enzymes, both native and recombinant, that catalyze the conversion of glycolaldehyde to glycolate and then to glyoxylate, which can enter the citric acid cycle via the action of malate synthase. Evidence is also presented that similar enzymes for this pentose sugar pathway are present in Sulfolobus acidocaldarius, and metabolic tracer studies in this archaeon demonstrate its in vivo operation in parallel with a route involving no aldol cleavage of the 2-keto-3-deoxy-pentanoates but direct conversion to the citric acid cycle C5-metabolite, 2-oxoglutarate.     

植物蛋白磷酸酶2C(PP2C)及其在信号转导中的作用

蛋白磷酸酶(protein phosphatase,PP)是蛋白质可逆磷酸化调节机制中的关键酶,蛋白磷酸酶2C(PP2C)是蛋白磷酸酶的一个分支。文章介绍了PP2C的结构及其在信号转导中的研究进展。     

Effect of Energy Metabolism on Protein Motility in the Bacterial Outer Membrane

We demonstrate the energy dependence of the motion of a porin, the λ-receptor, in the outer membrane of living Escherichia coli by single molecule investigations. By poisoning the bacteria with arsenate and azide, the bacterial energy metabolism was stopped. The motility of individual λ-receptors significantly and rapidly decreased upon energy depletion. We suggest two different causes for the ceased motility upon comprised energy metabolism: One possible cause is that the cell uses energy to actively wiggle its proteins, this energy being one order-of-magnitude larger than thermal energy. Another possible cause is an induced change in the connection between the λ-receptor and the membrane structure, for instance by a stiffening of part of the membrane structure. Treatment of the cells with ampicillin, which directly targets the bacterial cell wall by inhibiting cross-linking of the peptidoglycan layer, had an effect similar to energy depletion and the motility of the λ-receptor significantly decreased. Since the λ-receptor is closely linked to the peptidoglycan layer, we propose that λ-receptor motility is directly coupled to the constant and dynamic energy-consuming reconstruction of the peptidoglycan layer. The result of this motion could be to facilitate transport of maltose-dextrins through the porin.     

Cordycepin Inhibits Protein Synthesis and Cell Adhesion through Effects on Signal Transduction

3′-Deoxyadenosine, also known as cordycepin, is a known polyadenylation inhibitor with a large spectrum of biological activities, including anti-proliferative, pro-apoptotic and anti-inflammatory effects. In this study we confirm that cordycepin reduces the length of poly(A) tails, with some mRNAs being much more sensitive than others. The low doses of cordycepin that cause poly(A) changes also reduce the proliferation of NIH3T3 fibroblasts. At higher doses of the drug we observed inhibition of cell attachment and a reduction of focal adhesions. Furthermore, we observed a strong inhibition of total protein synthesis that correlates with an inhibition of mammalian target of rapamycin (mTOR) signaling, as observed by reductions in Akt kinase and 4E-binding protein (4EBP) phosphorylation. In 4EBP knock-out cells, the effect of cordycepin on translation is strongly reduced, confirming the role of this modification. In addition, the AMP-activated kinase (AMPK) was shown to be activated. Inhibition of AMPK prevented translation repression by cordycepin and abolished 4EBP1 dephosphorylation, indicating that the effect of cordycepin on mTOR signaling and protein synthesis is mediated by AMPK activation. We conclude that many of the reported biological effects of cordycepin are likely to be due to its effects on mTOR and AMPK signaling.     

Scapinin,the Protein Phosphatase 1 Binding Protein,Enhances Cell Spreading and Motility by Interacting with the Actin Cytoskeleton

Scapinin, also named phactr3, is an actin and protein phosphatase 1 (PP1) binding protein, which is expressed in the adult brain and some tumor cells. At present, the role(s) of scapinin in the brain and tumors are poorly understood. We show that the RPEL-repeat domain of scapinin, which is responsible for its direct interaction with actin, inhibits actin polymerization in vitro. Next, we established a Hela cell line, where scapinin expression was induced by tetracycline. In these cells, expression of scapinin stimulated cell spreading and motility. Scapinin was colocalized with actin at the edge of spreading cells. To explore the roles of the RPEL-repeat and PP1-binding domains, we expressed wild-type and mutant scapinins as fusion proteins with green fluorescence protein (GFP) in Cos7 cells. Expression of GFP-scapinin (wild type) also stimulated cell spreading, but mutation in the RPEL-repeat domain abolished both the actin binding and the cell spreading activity. PP1-binding deficient mutants strongly induced cell retraction. Long and branched cytoplasmic processes were developed during the cell retraction. These results suggest that scapinin enhances cell spreading and motility through direct interaction with actin and that PP1 plays a regulatory role in scapinin-induced morphological changes.     

植物细胞活性氧种类、代谢及其信号转导

越来越明显的证据表明,植物体十分活跃的产生着活性氧并将之作为信号分子、进而控制着诸如细胞程序性死亡、非生物胁迫响应、病原体防御和系统信号等生命过程,而不仅是传统意义上的活性氧是有氧代谢的附产物。日益增多的证据显示,由脱落酸、水杨酸、茉莉酸与乙烯以及活性氧所调节的激素信号途径,在生物和非生物胁迫信号的“交谈”中起重要作用。活性氧最初被认为是动物吞噬细胞在宿主防御反应时所释放的附产物,现在的研究清楚的表明,活性氧在动物和植物细胞信号途径中均起作用。活性氧可以诱导细胞程序性死亡或坏死、可以诱导或抑制许多基因的表达,也可以激活上述级联信号。近来生物化学与遗传学研究证实过氧化氢是介导植物生物胁迫与非生物胁迫的信号分子,过氧化氢的合成与作用似乎与一氧化氮有关系。过氧化氢所调节的下游信号包括钙“动员”、蛋白磷酸化和基因表达等。     

生物和非生物胁迫下的植物细胞中丝裂原活化蛋白激酶(MAPK)信号转导

介绍了生物和非生物胁迫下植物细胞中MAPK信号转导的研究进展。     

酪氨酸蛋白磷酸酶(PTPase)参与水杨酸调控蚕豆气孔运动的信号转导

研究酪氨酸蛋白磷酸酶(PTPase)的抑制剂氧化苯胂(PAO)、钒酸钠(NaVO3)和Zn2 对水杨酸(SA)调控蚕豆气孔运动影响的结果表明,0~1mmol·L-1 PAO、0~4mmol·L-1 NaVO3和0~4mmol·L-1Zn2 对光诱导蚕豆气孔开度变化的影响不大,但都可以抑制黑暗或SA诱导的气孔关闭,据此推测,PTPase可能参与SA诱导气孔关闭的信号转导过程。     

细胞分裂素:代谢、信号转导、交叉反应与农艺性状改良

在高等植物中,细胞分裂素通过对细胞分裂与分化的调节而广泛参与了对植物生长发育的调控。在过去的10余年,利用模式植物拟南芥的研究,在阐明细胞分裂素的代谢、转运与信号转导等方面取得了重要的进展。同时,关于细胞分裂素与其它信号途径之间存在的广泛交叉反应也受到了人们的注意。根据我们现有的知识,细胞分裂素信号转导是通过磷酸基团在一个双元组分系统之间的系列传递而完成的,该过程被称之为“磷酸接力传递”（phosphorelay）。细胞分裂素与其它信号途径的互作可能也主要是通过双元组分系统链接的。双元组分系统中目前已知的主要信号元件不仅表现出功能冗余性,同时在调控特定的植物生长发育过程时也具有特异性。本文在对细胞分裂素的代谢与转运过程简要评述的基础上,对其信号转导以及与其它信号途径间交叉反应的研究进展进行重点讨论,并展望细胞分裂素研究对重要农业性状改良的意义。     

COP9信号复合体:信号转导与蛋白质降解的接合器

COP9信号复合体(CSN)是细胞内高度保守的多亚基蛋白质复合物,主要定位于真核细胞的细胞核。在结构上与26S蛋白酶体“盖子”亚复合物高度相关。CSN的具体功能目前尚未明确,主要体现为两方面的活性：脱Nedd化作用和磷酸化作用;在细胞内同SCF遍在蛋白连接酶复合体等许多蛋白质复合物发生相互作用;调节多种信号分子靶向遍在蛋白-26S蛋白酶体的稳定性。因此,CSN是连接信号转导与蛋白质降解的分子平台。     

丙型肝炎病毒蛋白作用于细胞信号转导途径的研究进展

细胞信号转导异常往往与人类疾病的发生、发展密切相关。一些病毒致病和感染机制即为病毒抗原蛋白作用宿主细胞信号转导途径,导致宿主细胞内信号转导发生紊乱。丙型肝炎病毒（HCV）是引发慢性丙型肝炎,导致肝硬化和肝细胞癌发生的主要病原体,但目前HCV的致病机制与宿主内持续感染机制尚不清楚。HCV致病机制可能与HCV表达的蛋白质干扰宿主细胞信号转导途径而导致异常的细胞信号转导有关。研究HCV蛋白对宿主细胞信号转导途径的影响不仅有助于阐明其致病机制,还能为新药设计和寻找新的治疗方法提供新思路和新靶点。本文主要综述了近年来国内外有关HCV蛋白作用细胞信号转导途径的研究进展。     

细胞分裂素: 代谢、信号转导、交叉反应与农艺性状改良

在高等植物中, 细胞分裂素通过对细胞分裂与分化的调节而广泛参与了对植物生长发育的调控。在过去的10余年, 利用模式植物拟南芥的研究, 在阐明细胞分裂素的代谢、转运与信号转导等方面取得了重要的进展。同时, 关于细胞分裂素与其它信号途径之间存在的广泛交叉反应也受到了人们的注意。根据我们现有的知识, 细胞分裂素信号转导是通过磷酸基团在一个双元组分系统之间的系列传递而完成的, 该过程被称之为“磷酸接力传递”(phosphorelay)。细胞分裂素与其它信号途径的互作可能也主要是通过双元组分系统链接的。双元组分系统中目前已知的主要信号元件不仅表现出功能冗余性, 同时在调控特定的植物生长发育过程时也具有特异性。本文在对细胞分裂素的代谢与转运过程简要评述的基础上,对其信号转导以及与其它信号途径间交叉反应的研究进展进行重点讨论, 并展望细胞分裂素研究对重要农业性状改良的意义。     

芸苔属自交不亲和细胞信号转导的研究进展

在芸苔属植物的自交不亲和细胞信号转导过程中,信号分子-SCR配体是由花粉粒产生的,被柱头乳突细胞SRK受体识别后,进行细胞内信号转导.这对受体-配体是两个由S位点编码的且高度多态的蛋白质,它们决定着自交不亲和反应.配体是位于花粉粒表面的一个小的胞被蛋白,由SCR基因编码;受体是位于柱头乳突细胞原生质膜上的跨膜的蛋白质激酶,由SRK基因编码.在自交授粉过程中,配体SCR和受体SRK的相互作用激活了受体SRK,被激活的SRK通过其下游组分ARC1介导底物的泛肽化,然后泛肽化的底物在蛋白酶体/CSN中被降解,从而导致了自交不亲和性反应.这些降解的底物可能是促进花粉水合、萌发和花粉管生长的雌蕊亲和因子.主要针对芸苔属自交不亲和细胞信号转导作一综述.     

芸苔属自交不亲和细胞信号转导的研究进展

在芸苔属植物的自交不亲和细胞信号转导过程中,信号分子-SCR配体是由花粉粒产生的,被柱头乳突细胞SRK受体识别后,进行细胞内信号转导。这对受体-配体是两个由S位点编码的且高度多态的蛋白质,它们决定着自交不亲和反应。配体是位于花粉粒表面的一个小的胞被蛋白,由SCR基因编码;受体是位于柱头乳突细胞原生质膜上的跨膜的蛋白质激酶,由SRK基因编码。在自交授粉过程中,配体SCR和受体SRK的相互作用激活了受体SRK,被激活的SRK通过其下游组分ARC1介导底物的泛肽化,然后泛肽化的底物在蛋白酶体/CSN中被降解,从而导致了自交不亲和性反应。这些降解的底物可能是促进花粉水合、萌发和花粉管生长的雌蕊亲和因子。主要针对芸苔属自交不亲和细胞信号转导作一综述。     

脑信号蛋白与细胞运动及肿瘤

脑信号蛋白（semaphorin）是分泌的或膜相关糖蛋白,其通过与相应的受体结合后刺激激酶、调节RhoGTP酶,通过调节R-Ras调控整合素、细胞骨架,从而调控细胞运动。脑信号蛋白信号系统也调节肿瘤细胞的运动,调控肿瘤血管生成,并和肝细胞生长因子HGF/Met相偶联,控制肿瘤的侵袭转移。     

Ghrelin: Impact on Muscle Energy Metabolism in Sepsis

We investigated the effects of exogenous ghrelin on energy levels and tissue histology in skeletal muscle in experimentally lipopolysaccharide (LPS) induced septic rats. Male Wistar albino rats 200–250 g were separated into four groups; Control, LPS (5 mg/kg), Ghrelin (10 nmol/kg i.v.), and ghrelin+LPS. Gastrocnemius muscle tissue was taken and stained using modified Gomori trichrome (MGT), succinic dehydrogenase (SDH), and cytochrome oxidase (COX) and hematoxylin and eosin. In stained sections, histological score value was calculated according to the intensity and the distribution for MGT, SDH and COX stainings. Creatine, creatine phosphate, adenosine triphosphate (ATP), adenosine monophosphate (AMP) levels, and the ratios of AMP/ATP and CreaP/ATP were investigated using high performance liquid chromatography (HPLC) in muscle tissue. Significances between experimental groups were calculated with an analysis of variance (ANOVA) followed by Tukey’s tests. Myopathic changes were seen in the 50% of rats in the LPS group as rounding of muscle fibers and fiber size variation. In the ghrelin+LPS group, ghrelin treatment was reduced damage in skeletal muscle structure. There was no change in creatine or AMP levels between the groups. Ghrelin treatment significantly increased ATP values (P?<?0.01) and improved tissue histology in septic rats. Ratios of both AMP/ATP and CreaP/ATP were found increased in the septic group, but there were decreaments in both the ghrelin and ghrelin-treated septic groups. Ghrelin could play an important role in energy balance and muscle morphology in skeletal muscle during sepsis.     

Regulation of the Src Kinase-associated Phosphoprotein 55 Homologue by the Protein Tyrosine Phosphatase PTP-PEST in the Control of Cell Motility

PTP-PEST is a cytosolic ubiquitous protein tyrosine phosphatase (PTP) that contains, in addition to its catalytic domain, several protein-protein interaction domains that allow it to interface with several signaling pathways. Among others, PTP-PEST is a key regulator of cellular motility and cytoskeleton dynamics. The complexity of the PTP-PEST interactome underscores the necessity to identify its interacting partners and physiological substrates in order to further understand its role in focal adhesion complex turnover and actin organization. Using a modified yeast substrate trapping two-hybrid system, we identified a cytosolic adaptor protein named Src kinase-associated phosphoprotein 55 homologue (SKAP-Hom) as a novel substrate of PTP-PEST. To confirm PTP-PEST interaction with SKAP-Hom, in vitro pull down assays were performed demonstrating that the PTP catalytic domain and Proline-rich 1 (P1) domain are respectively binding to the SKAP-Hom Y260 and Y297 residues and its SH3 domain. Subsequently, we generated and rescued SKAP-Hom-deficient mouse embryonic fibroblasts (MEFs) with WT SKAP-Hom, SKAP-Hom tyrosine mutants (Y260F, Y260F/Y297F), or SKAP-Hom SH3 domain mutant (W335K). Given the role of PTP-PEST, wound-healing and trans-well migration assays were performed using the generated lines. Indeed, SKAP-Hom-deficient MEFs showed a defect in migration compared with WT-rescued MEFs. Interestingly, the SH3 domain mutant-rescued MEFs showed an enhanced cell migration corresponding potentially with higher tyrosine phosphorylation levels of SKAP-Hom. These findings suggest a novel role of SKAP-Hom and its phosphorylation in the regulation of cellular motility. Moreover, these results open new avenues by which PTP-PEST regulates cellular migration, a hallmark of metastasis.     

绿色荧光蛋白的特性及其在信号转导中的应用

细胞和分子生物学的最终目标是 ,明确细胞内的各种事件是如何发生的 ,明了细胞内复杂的动态变化的生化机制。绿色荧光蛋白 (ｇｒｅｅｎｆｌｕｏｒｅｓｃｅｎｔｐｒｏｔｅｉｎ ,ＧＦＰ)自从克隆、表达之后 ,以其良好的物理特性及荧光特性而成为良好的报告基因和荧光标记分子 ,并在探索生命现象过程中得到了非常广泛的应用。ＧＦＰ作为报告基因 ,可用在活细胞中直接观察蛋白质向细胞器 ,如细胞核、内质网中运动 ;作为荧光标记分子 ,ＧＦＰ既具有敏感的标记检测率 ,又没有放射性的危害 ;最近又发现ＧＦＰ是一个良好的细胞间信号传递的动态标记…     

G蛋白及其在细胞信号转导中的作用

概述G蛋白的结构、种类、转导信息的机制,细菌毒素对G蛋白调节的影响及研究G蛋的意义和今后发展的重点。     

Stochastic Simulation of Signal Transduction: Impact of the Cellular Architecture on Diffusion

The transduction of signals depends on the translocation of signaling molecules to specific targets. Undirected diffusion processes play a key role in the bridging of spaces between different cellular compartments. The diffusion of the molecules is, in turn, governed by the intracellular architecture. Molecular crowding and the cytoskeleton decrease macroscopic diffusion. This article shows the use of a stochastic simulation method to study the effects of the cytoskeleton structure on the mobility of macromolecules. Brownian dynamics and single particle tracking were used to simulate the diffusion process of individual molecules through a model cytoskeleton. The resulting average effective diffusion is in line with data obtained in the in vitro and in vivo experiments. It shows that the cytoskeleton structure strongly influences the diffusion of macromolecules. The simulation method used also allows the inclusion of reactions in order to model complete signaling pathways in their spatio-temporal dynamics, taking into account the effects of the cellular architecture.