首页 | 本学科首页   官方微博 | 高级检索  
相似文献
 共查询到20条相似文献,搜索用时 46 毫秒
1.
Assembly of clathrin-coated pits onto purified plasma membranes   总被引:32,自引:0,他引:32  
During receptor-mediated endocytosis, coated pits invaginate to form coated vesicles, clathrin and associated proteins dissociate from the vesicle membrane, and these proteins form new coated pits at the cell surface. As a means of elucidating molecular mechanisms that govern the function of coated pits, the assembly phase of this cycle was reconstituted by incubating purified membranes that were treated to remove endogenous coated pits with cytoplasm extracted from cultured cells. The in vitro assembly of coated pits on these membranes satisfactorily mimics many features of coated pit formation in the intact cell. These studies indicate that: the membranes contain a limited number of coated pit assembly sites that bind clathrin with high affinity; the half-time for assembly is 5 minutes both at 4 degrees C and 37 degrees C; during assembly, proteins with molecular sizes of 180, 110, and 36 kilodaltons are recruited to the plasma membrane; and assembly is not dependent on adenosine triphosphate, but this nucleotide triggers a temperature-dependent loss of coated pits that are assembled in the absence of adenosine triphosphate.  相似文献   

2.
Living with clathrin: its role in intracellular membrane traffic   总被引:53,自引:0,他引:53  
Clathrin polymerization at the cytoplasmic side of the plasma membrane forms coated pits and vesicles that mediate uptake of cell surface receptors. Clathrin-coated vesicles have also been implicated in protein export but definition of their precise role has been controversial. Recent advances in characterization of the clathrin subunits and additional coated vesicle components have identified molecular interactions involved in clathrin polymerization and coated vesicle formation, and have provided new approaches to investigating its function. These studies suggest that clathrin's role, in both inward and outward membrane traffic, is to facilitate receptor transport by a concentration and sorting process that initiates targeting to specific intracellular compartments.  相似文献   

3.
对禾谷缢管蚜和麦长管蚜传播小麦矮病毒GPV株系的超微结构观察发现:禾谷管蚜取食8h时,病毒进入蚜虫后肠肠腔中,12-24h后进入后肠细胞质,GPV与肠内壁顶端细胞膜接触,接触部位细胞膜逐渐下陷,形成小穴,向内凹陷发育为管状泡囊,而后进入血淋巴液,当病毒被附唾液腺吸收后蚜虫将病毒排吐进入唾液管中。  相似文献   

4.
Transport between compartments of eukaryotic cells is mediated by coated vesicles. The archetypal protein coats COPI, COPII, and clathrin are conserved from yeast to human. Structural studies of COPII and clathrin coats assembled in vitro without membranes suggest that coat components assemble regular cages with the same set of interactions between components. Detailed three-dimensional structures of coated membrane vesicles have not been obtained. Here, we solved the structures of individual COPI-coated membrane vesicles by cryoelectron tomography and subtomogram averaging of in vitro reconstituted budding reactions. The coat protein complex, coatomer, was observed to adopt alternative conformations to change the number of other coatomers with which it interacts and to form vesicles with variable sizes and shapes. This represents a fundamentally different basis for vesicle coat assembly.  相似文献   

5.
分析氟涂层刀具铣削饰面刨花板的切削性能,为氟涂层刀具在木材切削加工领域应用提供理论指导。利用CNC加工中心分别进行未涂层刀具和氟涂层刀具铣削饰面刨花板试验,采用工具显微镜和扫描电镜拍摄刀具前后刀面的磨损形貌,采用超景深三维显微镜对饰面刨花板的切削加工表面进行粗糙度测量,研究氟涂层刀具铣削饰面刨花板的切削性能。结果表明,在相同的铣削长度条件下,氟涂层刀具铣削饰面刨花板的后刀面磨损带宽度显著低于未涂层刀具的后刀面磨损带宽度,且随着铣削长度的逐渐增加,未涂层刀具磨损带宽度增加急剧,而涂层刀具磨损带宽度增大缓慢;氟涂层刀具主要为磨料磨损,可以有效减少胶黏剂和刨花颗粒产生的黏结磨损,且降低崩刃现象;在相同的铣削长度条件下,氟涂层刀具铣削饰面刨花板的切削表面粗糙度值明显低于未涂层刀具铣削饰面刨花板的切削表面粗糙度值,且铣削饰面刨花板的刨花层可显著减少表面毛刺和凹坑,且饰面耐磨层崩边较少。因此,氟涂层刀具铣削饰面刨花板可以有效降低刀具磨损,提高饰面刨花板切削表面质量。  相似文献   

6.
质膜ATP酶介导机械伤害诱导豌豆叶片的防御反应   总被引:1,自引:0,他引:1  
刘艳  黄卫东  陈贵林  岳鑫 《中国农业科学》2010,43(16):3411-3417
【目的】研究质膜ATP酶(H+-ATPase)在机械胁迫诱导植物防御反应形成中的作用。【方法】通过酶活性生化测定、Western blot检测和电镜细胞化学观测方法,研究机械伤害对豌豆幼苗质膜H+-ATPase活性的影响;同时结合药理学实验,进一步探讨了质膜H+-ATPase与活性氧积累在伤害防御反应中的关系。【结果】机械胁迫能诱导质膜H+-ATPase活性提高,但并未改变该酶蛋白含量;抑制质膜H+-ATPase活性,伤诱导的H2O2累积和苯丙氨酸解氨酶活性提高随之被抑制,活性氧清除剂处理对伤诱导质膜H+-ATPase活性提高没有影响。【结论】质膜H+-ATPase参与机械胁迫诱导防御反应相关的信号传递,该酶可能作用于活性氧信号上游,并通过蛋白磷酸化调节来诱导下游防御反应形成。  相似文献   

7.
• Herbivory and mechanical wounding elicited electrical signals.• Petiole wounding elicited stronger electrical signals than did leaflet wounding.• Leaflet wounding elicited electrical signals and JA signaling within a compound leaf.• GLR3.3 and GLR3.5 mediated leaflet-to-leaflet electrical signal transduction.• JA synthesis and Helicoverpa armigera resistance were reduced in glr3.3/3.5 plants.Electrical signals commonly occur in plants in response to various environmental changes and have a dominant function in plant acclimation. The transduction of wound-elicited electrical signals in the model plant species Arabidopsis has been characterized but the characteristics of electrical signal transduction in response to herbivory or wounding in crop species remain unknown. Here, the features of electrical signals elicited by insect herbivory and wounding in tomato were investigated. Unlike those in Arabidopsis, wounding tomato leaves did not cause leaf-to-leaf electrical signal transduction. In contrast, electrical signals elicited in response to petiole wounding were stronger and more strongly transduced. Leaflet wounding also activated electrical signal transduction and jasmonic acid (JA) signaling within the whole compound leaf. It was also demonstrated that tomato glutamate receptor-like 3.3 (GLR3.3) and GLR3.5 mediated leaflet-to-leaflet electrical signal transduction. Herbivory-induced JA accumulation and Helicoverpa armigera resistance were reduced in glr3.3/3.5 plants. This work reveals the nature of electrical signal transduction in tomato and emphasizes the key roles of GLR3.3 and GLR3.5 in electrical signal transduction and JA signaling activation.  相似文献   

8.
The surface membrane of cells is studded with morphologically distinct regions, or domains, like microvilli, cell-cell junctions, and coated pits. Each of these domains is specialized for a particular function, such as nutrient absorption, cell-cell communication, and endocytosis. Lipid domains, which include caveolae and rafts, are one of the least understood membrane domains. These domains are high in cholesterol and sphingolipids, have a light buoyant density, and function in both endocytosis and cell signaling. A major mystery, however, is how resident molecules are targeted to lipid domains. Here, we propose that the molecular address for proteins targeted to lipid domains is a lipid shell.  相似文献   

9.
The roles of on endogenous jasmonates (JAs) and salicylic acid (SA) in wounding response were investigated. Pea (Pisum sativum L.) seedlings were treated with three different methods including mechanical wounding, JAs application, and SA application. The contents of endogenous JAs and SA, as well as the activities of the related enzymes were detected by enzyme-linked immunosorbent assay (ELISA), high performance liquid chromatography (HPLC), and spectrophotometer, respectively. The results showed that endogenous JA rapidly accumulated within 30 min after wounding. The increase in the activities of both lipoxygenase (LOX) and allene oxide synthase (AOS) lagged behind JAs burst. A second slight increase in JAs level was observed at 24 h after wounding treatment, and at the same time point, higher activities of LOX and AOS were also detected. Endogenous free SA content decreased accompanied with JAs burst. Effects of exogenous JA application were similar to those of wounding treatment on endogenous SA level and phenylalanine ammonia lyase (PAL) activity, whereas exogenous SA application led to the significant inhibition of LOX and AOS activities and the decrease of endogenous JAs level at the early stage of treatment. It is thus suggested that JAs burst and SA decrease in early response to wounding may constitute an important mechanism by which plant starts the related defense reaction and adapts to wounding stress.  相似文献   

10.
豌豆叶片内源水杨酸和茉莉酸类物质对机械伤害的响应   总被引:3,自引:0,他引:3  
 【目的】研究内源茉莉酸类(JAs)和水杨酸(SA)对机械伤害的响应特点。【方法】以豌豆幼苗为试材,设置机械伤害、外施茉莉酸(JA)和SA 3个处理,分别采用ELISA、HPLC和分光光度法测定内源JAs与SA含量,及其合成关键酶活性。【结果】伤害处理后30 min,内源JAs含量显著增加,随后迅速下降;JAs生物合成关键酶脂氧合酶(LOX)和丙二烯氧化物合成酶(AOS)活性增加滞后于JAs含量增加。伤害处理后24 h,内源JAs含量又有小幅升高,此时LOX和AOS活性也维持在较高水平。与此相对应,内源自由态SA在JAs突发时呈下降趋势。外源JA处理后内源SA和苯丙氨酸解氨酶(PAL)活性变化同伤害处理相似。外源SA处理初期显著抑制了LOX和AOS活性,同时叶片内源JAs含量降低。【结论】伤害早期内源JAs含量升高和SA含量降低是植物启动相应防御反应和抵御伤害胁迫的重要机制。  相似文献   

11.
The molecular size of the plasma LDL (low density lipoprotein) receptor synthesized by cultured fibroblasts from a patient with the internalization-defective form of familial hypercholesterolemia (FH 274) was smaller by 10,000 daltons than the size of the normal LDL receptor. The segment of the gene encoding the truncated portion of the FH 274 receptor was cloned into bacteriophage lambda. Comparison of the nucleotide sequences of the normal and FH 274 genes revealed a 5-kilobase deletion, which eliminated the exons encoding the membrane-spanning region and the carboxyl terminal cytoplasmic domain of the receptor. The deletion appeared to be caused by a novel intrastrand recombination between two repetitive sequences of the Alu family that were oriented in opposite directions. The truncated receptors lack membrane-spanning regions and cytoplasmic domains; they are largely secreted into the culture medium, but a small fraction remains adherent to the cell surface. The surface-adherent receptors bind LDL, but they are unable to cluster in coated pits, thus explaining the internalization-defective phenotype.  相似文献   

12.
通过实时荧光定量PCR(qRealTime-PCR)检测柑橘伤害诱导基因(CsWIP1)在乙烯胁迫和机械伤害过程中的表达情况.结果显示:CsWIP1仅在外源乙烯处理的叶片和果实离层(AZ-C)中表达,而在伤害、1-MCP+乙烯、1-MCP+伤害处理的叶片中均不表达,由此可见CsWIP1仅受到乙烯信号传导途径调控.  相似文献   

13.
不同生境下蓝桉木材扫描电镜观察的研究   总被引:2,自引:0,他引:2  
用扫描电镜对不同生境下的蓝桉(EucalyptusglobulusLabill.)木材结构进行了解剖学观察,不同生境下的蓝桉扫描结构表明,导管壁上具A,B两种类型的附物纹孔,但以A型为主,也具具缘纹孔、联合状纹孔和火山状纹孔;管胞壁上具附物纹孔,但附物较少;纤维管胞壁上具具缘纹孔和单纹孔;纤维壁上具长椭圆形的单纹孔;射线细胞内具晶体和晶体类似物。并分析了附物纹孔的大小与生态因子之间的关系。  相似文献   

14.
The Golgi-localized, gamma-ear-containing, adenosine diphosphate ribosylation factor-binding proteins (GGAs) are multidomain proteins that bind mannose 6-phosphate receptors (MPRs) in the Golgi and have an essential role in lysosomal enzyme sorting. Here the GGAs and the coat protein adaptor protein-1 (AP-1) were shown to colocalize in clathrin-coated buds of the trans-Golgi networks of mouse L cells and human HeLa cells. Binding studies revealed a direct interaction between the hinge domains of the GGAs and the gamma-ear domain of AP-1. Further, AP-1 contained bound casein kinase-2 that phosphorylated GGA1 and GGA3, thereby causing autoinhibition. This could induce the directed transfer of the MPRs from GGAs to AP-1. MPRs that are defective in binding to GGAs are poorly incorporated into AP-1-containing clathrin-coated vesicles. Thus, the GGAs and AP-1 interact to package MPRs into AP-1-containing coated vesicles.  相似文献   

15.
试验结果表明,葡萄植株经过不同伤害处理后,所诱导的挥发物组成存在明显的不同,机械损伤后葡萄植株释放的挥发物主要是醇、醛和酯类物质,二点叶螨取食后释放出的挥发物以萜烯类化合物为主。  相似文献   

16.
以茄子为供试材料,研究机械损伤、二点叶螨取食对茄子损伤诱导挥发物的影响,结果表明:机械损伤后,茄子植株释放的挥发物主要是绿叶性气体;遭受二点叶螨取食后释放出的挥发物与机械损伤植株存在很大不同,许多新的组分被诱导出来。  相似文献   

17.
There is a series of pits in the scales of the rostrum and posterior portion of the lower lips in some pythons and boas. In the Australian python Morelia spilotes, these pits are innervated by the maxillary and mandibular branches of the trigeminal nerve. Structural and neurophysiological evidence indicate that in the pits there are receptors that function as detectors of radiant heat flux.  相似文献   

18.
Stainless steels undergo a sharp rise in pitting corrosion rate as the potential, solution concentration, or temperature is changed only slightly. We report experiments using real-time microscopic in situ visualizations that resolve the nucleation and evolution of individual pits during the transition. They suggest that the sudden onset of corrosion is explained by an explosive autocatalytic growth in the number of metastable pits and that stabilization of individual pits takes place only later. This finding agrees with a theoretical approach treating the onset of pitting corrosion as a cooperative critical phenomenon resulting from interactions among metastable pits, and it extends perspectives on the control and prevention of corrosion onset.  相似文献   

19.
Damage to the vessel wall is a signal for endothelial migration and replication and for platelet release at the site of injury. Addition of transforming growth factor-beta (TGF-beta) purified from platelets to growing aortic endothelial cells inhibited [3H]thymidine incorporation in a concentration-dependent manner. A transient inhibition of DNA synthesis was also observed in response to wounding; cell migration and replication are inhibited during the first 24 hours after wounding. By 48 hours after wounding both TGF-beta-treated and -untreated cultures showed similar responses. Flow microfluorimetric analysis of cell cycle distribution indicated that after 24 hours of exposure to TGF-beta the cells were blocked from entering S phase, and the fraction of cells in G1 was increased. The inhibition of the initiation of regeneration by TGF-beta could allow time for recruitment of smooth muscle cells into the site of injury by other platelet components.  相似文献   

20.
Adaptor protein 180 (AP180) and its homolog, clathrin assembly lymphoid myeloid leukemia protein (CALM), are closely related proteins that play important roles in clathrin-mediated endocytosis. Here, we present the structure of the NH2-terminal domain of CALM bound to phosphatidylinositol-4,5- bisphosphate [PtdIns(4,5)P2] via a lysine-rich motif. This motif is found in other proteins predicted to have domains of similar structure (for example, Huntingtin interacting protein 1). The structure is in part similar to the epsin NH2-terminal (ENTH) domain, but epsin lacks the PtdIns(4,5)P2-binding site. Because AP180 could bind to PtdIns(4,5)P2 and clathrin simultaneously, it may serve to tether clathrin to the membrane. This was shown by using purified components and a budding assay on preformed lipid monolayers. In the presence of AP180, clathrin lattices formed on the monolayer. When AP2 was also present, coated pits were formed.  相似文献   

设为首页 | 免责声明 | 关于勤云 | 加入收藏

Copyright©北京勤云科技发展有限公司  京ICP备09084417号