全文获取类型
收费全文 | 731篇 |
免费 | 61篇 |
出版年
2023年 | 3篇 |
2022年 | 7篇 |
2021年 | 17篇 |
2020年 | 7篇 |
2019年 | 15篇 |
2018年 | 14篇 |
2017年 | 17篇 |
2016年 | 12篇 |
2015年 | 44篇 |
2014年 | 50篇 |
2013年 | 52篇 |
2012年 | 82篇 |
2011年 | 73篇 |
2010年 | 38篇 |
2009年 | 30篇 |
2008年 | 47篇 |
2007年 | 46篇 |
2006年 | 48篇 |
2005年 | 42篇 |
2004年 | 45篇 |
2003年 | 45篇 |
2002年 | 25篇 |
2001年 | 7篇 |
2000年 | 2篇 |
1999年 | 4篇 |
1998年 | 2篇 |
1997年 | 1篇 |
1996年 | 1篇 |
1995年 | 1篇 |
1994年 | 1篇 |
1992年 | 3篇 |
1991年 | 2篇 |
1990年 | 2篇 |
1989年 | 2篇 |
1988年 | 2篇 |
1986年 | 1篇 |
1981年 | 1篇 |
1975年 | 1篇 |
排序方式: 共有792条查询结果,搜索用时 31 毫秒
51.
52.
Andrea Erlbruch Chien‐Wen Hung Joerg Seidler Katrin Borrmann Frank Gesellchen Norbert König Dieter Kübler Friedrich W. Herberg Wolf D. Lehmann Dirk Bossemeyer 《Proteomics》2010,10(16):2890-2900
An expression‐uncoupled tandem affinity purification assay is introduced which differs from the standard TAP assay by uncoupling the expression of the TAP‐bait protein from the target cells. Here, the TAP‐tagged bait protein is expressed in Escherichia coli and purified. The two concatenated purification steps of the classical TAP are performed after addition of the purified bait to brain tissue homogenates, cell and nuclear extracts. Without prior genetic manipulation of the target, upscaling, free choice of cell compartments and avoidance of expression triggered heat shock responses could be achieved in one go. By the strategy of separating bait expression from the prey protein environment numerous established, mostly tissue‐specific binding partners of the protein kinase A catalytic subunit Cβ1 were identified, including interactions in binary, ternary and quaternary complexes. In addition, the previously unknown small molecule inhibitor‐dependent interaction of Cβ1 with the cell cycle and apoptosis regulatory protein‐1 was verified. The uncoupled tandem affinity purification procedure presented here expands the application range of the in vivo TAP assay and may serve as a simple strategy for identifying cell‐ and tissue‐specific protein complexes. 相似文献
53.
Joerg Flemmig 《Journal of inorganic biochemistry》2010,104(7):759-764
The close association of the heme enzyme myeloperoxidase to phosphatidylserine epitopes on the surface of non-vital polymorphonuclear leukocytes (PMNs) and other apoptotic cells at inflammatory sites favours modifications of this phospholipid by myeloperoxidase products. As detected by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry, ammonium ions inhibit in a concentration-dependent manner the hypochlorous acid-mediated formation of aldehyde and nitrile products from 1,2-dipalmitoyl-sn-glycero-3-phosphoserine (DPPS). Concomitantly, the formation of monochloramine (NH2Cl) raises with increasing NH4+ concentrations. A transchlorination from monochlorinated O-phospho-l-serine to NH4+ with the formation of NH2Cl occurs only when extraordinary high NH4+ concentrations are applied. Due to the low rate of 0.044 M− 1 s− 1 for this process, a transhalogenation reaction from transient chlorinated intermediates of the serine moiety to NH4+ can be ruled out as an important process contributing to the HOCl-mediated formation of NH2Cl. A significant formation of NH2Cl by myeloperoxidase interacting with DPPS in the presence of ammonium ions takes only place at acidic pH values around 5, a scenario that may occur in phagosomes of macrophages after the uptake of apoptotic PMNs. 相似文献
54.
Andrea Dlasková Tomáš Špaček Jitka Šantorová Lydie Plecitá-Hlavatá Zuzana Berková František Saudek Mark Lessard Joerg Bewersdorf Petr Ježek 《BBA》2010,1797(6-7):1327-1341
Insulin production in pancreatic β-cells is critically linked to mitochondrial oxidative phosphorylation. Increased ATP production triggered by blood glucose represents the β-cells' glucose sensor. Type-2 diabetes mellitus results from insulin resistance in peripheral tissues and impaired insulin secretion. Pathology of diabetic β-cells might be reflected by the altered morphology of mitochondrial network. Its characterization is however hampered by the complexity and density of the three-dimensional (3D) mitochondrial tubular networks in these cell types. Conventional confocal microscopy does not provide sufficient axial resolution to reveal the required details; electron tomography reconstruction of these dense networks is still difficult and time consuming. However, mitochondrial network morphology in fixed cells can also be studied by 4Pi microscopy, a laser scanning microscopy technique which provides an ~ 7-fold improved axial resolution (~ 100 nm) over conventional confocal microscopy. Here we present a quantitative study of these networks in insulinoma INS-1E cells and primary β-cells in Langerhans islets. The former were a stably-transfected cell line while the latter were transfected with lentivirus, both expressing mitochondrial matrix targeted redox-sensitive GFP. The mitochondrial networks and their partial disintegration and fragmentation are revealed by carefully created iso-surface plots and their quantitative analysis. We demonstrate that β-cells within the Langerhans islets from diabetic Goto Kakizaki rats exhibited a more disintegrated mitochondrial network compared to those from control Wistar rats and model insulinoma INS-1E cells. Standardization of these patterns may lead to development of morphological diagnostics for Langerhans islets, for the assessment of β-cell condition, before their transplantations. 相似文献
55.
Bräutigam A Bomke S Pfeifer T Karst U Krauss GJ Wesenberg D 《Metallomics : integrated biometal science》2010,2(8):565-570
A method for the identification and quantification of canonic and isoforms of phytochelatins (PCs) from Chlamydomonas reinhardtii was developed. After disulfide reduction with tris(2-carboxyethyl)phosphine (TCEP) PCs were derivatized with ferrocenecarboxylic acid (2-maleimidoyl)ethylamide (FMEA) in order to avoid oxidation of the free thiol functions during analysis. Liquid chromatography (LC) coupled to electrospray mass spectrometry (ESI-MS) and inductively coupled plasma-mass spectrometry (ICP-MS) was used for rapid and quantitative analysis of the precolumn derivatized PCs. PC(2-4), CysGSH, CysPC(2-4), CysPC(2)desGly, CysPC(2)Glu and CysPC(2)Ala were determined in the algal samples depending on the exposure of the cells to cadmium ions. 相似文献
56.
M. Cristina Palmieri Christian Lindermayr Hermann Bauwe Clara Steinhauser Joerg Durner 《Plant physiology》2010,152(3):1514-1528
Mitochondria play an essential role in nitric oxide (NO) signal transduction in plants. Using the biotin-switch method in conjunction with nano-liquid chromatography and mass spectrometry, we identified 11 candidate proteins that were S-nitrosylated and/or glutathionylated in mitochondria of Arabidopsis (Arabidopsis thaliana) leaves. These included glycine decarboxylase complex (GDC), a key enzyme of the photorespiratory C2 cycle in C3 plants. GDC activity was inhibited by S-nitrosoglutathione due to S-nitrosylation/S-glutathionylation of several cysteine residues. Gas-exchange measurements demonstrated that the bacterial elicitor harpin, a strong inducer of reactive oxygen species and NO, inhibits GDC activity. Furthermore, an inhibitor of GDC, aminoacetonitrile, was able to mimic mitochondrial depolarization, hydrogen peroxide production, and cell death in response to stress or harpin treatment of cultured Arabidopsis cells. These findings indicate that the mitochondrial photorespiratory system is involved in the regulation of NO signal transduction in Arabidopsis.Nitric oxide (NO) has emerged as a new chemical messenger in plant biology. It can interact with a variety of intracellular and extracellular targets, acting as either a cytotoxic or a cytoprotective agent. NO stimulates seed germination in different species, and a decrease in NO levels has been associated with fruit maturation and senescence of flowers (Beligni and Lamattina, 2001). NO production has been observed in response to several biotic and abiotic stimuli, such as pathogen infection, bacterial elicitors, high temperature, osmotic stress, and UV-B light (Durner et al., 1998; Barroso et al., 1999; Krause and Durner, 2004; Zeidler et al., 2004; Shapiro, 2005; Corpas et al., 2008; Kolbert et al., 2008; Zhao et al., 2009).Despite the proven importance of NO, little is known about signaling pathways downstream from it. During both programmed cell death and defense responses, NO requires cGMP and cADP Rib as secondary messengers (Wendehenne et al., 2001). Furthermore, NO activates mitogen-activated protein kinases in different plant species during stress signaling (Nakagami et al., 2005). However, direct biological activity of NO arises from chemical reactions between proteins and NO itself (Foster and Stamler, 2004; Dahm et al., 2006). S-Nitrosylation is a labile posttranslational modification with a half-life of seconds to a few minutes and represents a very sensitive mechanism for regulating cellular processes (Hess et al., 2005). More than 100 candidate S-nitrosylated proteins were identified from extracts of Arabidopsis (Arabidopsis thaliana) cultured cells treated with the NO donor S-nitrosoglutathione (GSNO) and from Arabidopsis leaves treated with gaseous NO (Lindermayr et al., 2005). Using the same proteomic approach, changes were characterized in S-nitrosylated proteins in Arabidopsis leaves undergoing a hypersensitive response (Romero-Puertas et al., 2008).In animals, mitochondria play a crucial role in S-nitrosylation-dependent NO signaling (Foster and Stamler, 2004). The mitochondrion is an essential organelle for normal cellular function, being an important site of ATP synthesis and an integrator for apoptotic signaling (Skulachev, 1999). Mitochondria interact with NO at several levels. One particularly well-characterized example is the inhibition of complex IV (cytochrome c oxidase) via binding of NO to its binuclear CuB/heme a3 site (Cleeter et al., 1994). There are several reasons why S-nitrosylation may be an important mitochondrial regulatory mechanism. For example, mitochondria contain sizeable pools of thiols and transition metals, all of which are known to modulate nitrosothiol (SNO) biochemistry (Foster and Stamler, 2004). In addition, mitochondria are highly membranous and accumulate lipophilic molecules such as NO. Interesting in this respect is the fact that the formation of the S-nitrosylating intermediate N2O3 is enhanced within membranes (Burwell et al., 2006).The role of mitochondria in stress-related responses has been investigated in both animals and plants. Endogenous nitrosylation of the catalytic Cys site of a subset of mitochondrial caspases serves as an on/off switch regulating caspase activity during apoptosis (Mannick et al., 2001). Moreover, cytochrome c, which is modified by NO at its heme iron during apoptosis, is released from mitochondria into the cytoplasm, which plays a critical role in many forms of apoptosis by stimulating apoptosome formation and subsequent caspase activation (Schonhoff et al., 2003). We previously showed that a prime target of NO in plants is the mitochondrial apparatus, causing an inhibition of KCN-sensitive respiration and an activation of alternative respiration via alternative oxidase (AOX; Huang et al., 2002; Krause and Durner, 2004; Livaja et al., 2008).The aim of this study was to identify possible targets for S-nitrosylation in mitochondria of Arabidopsis leaves in order to gain more insight into the regulatory function of NO at the protein level. Using a proteomic approach involving the highly specific biotin-switch method for detection and purification of S-nitrosylated proteins (Jaffrey and Snyder, 2001) in conjunction with liquid chromatography and tandem mass spectrometry (nanoLC/MS/MS), we could identify 11 mitochondrial proteins as targets for S-nitrosylation. Among these identified proteins, we focused our attention on the P-subunit of the Gly decarboxylase complex (GDC), which is an integral part of the photorespiratory system. Since the release of apoptotic factors from mitochondria may be a result of inhibition of respiration, transition of mitochondrial permeability, and formation of reactive oxygen species (ROS; Saviani et al., 2002; Taylor et al., 2004; Chen and Gibson, 2008), we investigated the molecular mechanism and the function of GDC-Cys modification in Arabidopsis. 相似文献
57.
DNA cleavage is a biochemical hallmark of apoptosis. In humans, apoptotic DNA cleavage is executed by DNA fragmentation factor (DFF) 40. In proliferating cells DFF40 is expressed in the presence of its chaperone and inhibitor DFF45, which results in the formation of the DFF complex. Here, we present a systematic analysis of the nuclear import of the DFF complex. Our in vitro experiments demonstrate that the importin alpha/beta-heterodimer mediates the translocation of the DFF complex from the cytoplasm to the nucleus. Both DFF subunits interact directly with the importin alpha/beta-heterodimer. However, importin alpha/beta binds more tightly to the DFF complex compared with the individual subunits. Additionally, the isolated C-terminal regions of both DFF subunits together bind importin alpha/beta more strongly than the individual C termini. Our results from in vivo studies reveal that the C-terminal regions of both DFF subunits harbor nuclear localization signals. Furthermore, nuclear import of the DFF complex requires the C-terminal regions of both subunits. In more detail, one basic cluster in the C-terminal region of each subunit, DFF40 (RLKRK) and DFF45 (KRAR), is essential for nuclear accumulation of the DFF complex. Based on these findings two alternative models for the interaction of importin alpha/beta with the DFF complex are presented. 相似文献
58.
Identification of genes involved in swarming motility using a Pseudomonas aeruginosa PAO1 mini-Tn5-lux mutant library
下载免费PDF全文
![点击此处可从《Journal of bacteriology》网站下载免费的PDF全文](/ch/ext_images/free.gif)
During a screening of a mini-Tn5-luxCDABE transposon mutant library of Pseudomonas aeruginosa PAO1 for alterations in swarming motility, 36 mutants were identified with Tn5 insertions in genes for the synthesis or function of flagellin and type IV pilus, in genes for the Xcp-related type II secretion system, and in regulatory, metabolic, chemosensory, and hypothetical genes with unknown functions. These mutants were differentially affected in swimming and twitching motility but in most cases had only a minor additional motility defect. Our data provide evidence that swarming is a more complex type of motility, since it is influenced by a large number of different genes in P. aeruginosa. Conversely, many of the swarming-negative mutants also showed an impairment in biofilm formation, indicating a strong relationship between these types of growth states. 相似文献
59.
Background: Complex procedures involving a facebow transfer and the use of lingualised teeth are deemed to have a positive influence on the chewing ability with complete dentures. Objectives: To determine if patients’ ratings of their ability to chew depend on the method of complete denture fabrication. Methods: Edentulous patients (n = 20) participated in a within‐subject crossover trial. Each patient received two sets of new complete dentures. One pair was manufactured based on intraoral tracing of centric relation and facebow transfer; semi‐anatomical teeth with lingualised occlusion denture (LOD) were chosen. The second pair was made using a simplified procedure without facebow transfer; jaw relations were recorded with wax occlusion rims, and anatomical teeth with a first premolar/canine‐guidance (CGD) were selected. The dentures were delivered in randomised order, and each was worn for 3 months. Three months after delivery, patients’ ratings of each new prosthesis were recorded on visual analogue scales for their ability to chew seven index foods. Repeated measurements analysis of variance was performed to investigate possible carry‐over effects accounting for confounding by treatment period. Results: When comparing the two treatments, participants rated their ability to chew in general, to masticate carrots, hard sausage, steak and raw apple in particular, was significantly better with the CGD (anatomical teeth) than with the LOD (p < 0.05). Conclusion: Comprehensive methods for the fabrication of complete dentures including semi‐anatomical lingualised teeth and a full registration do not seem to influence the perceived chewing ability, when compared with more simple procedures. Chewing ability for tough foods appears to benefit from the use of anatomical teeth. 相似文献
60.