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61.
62.
Structure determination of T cell protein-tyrosine phosphatase 总被引:2,自引:0,他引:2
Iversen LF Moller KB Pedersen AK Peters GH Petersen AS Andersen HS Branner S Mortensen SB Moller NP 《The Journal of biological chemistry》2002,277(22):19982-19990
Protein-tyrosine phosphatase 1B (PTP1B) has recently received much attention as a potential drug target in type 2 diabetes. This has in particular been spurred by the finding that PTP1B knockout mice show increased insulin sensitivity and resistance to diet-induced obesity. Surprisingly, the highly homologous T cell protein-tyrosine phosphatase (TC-PTP) has received much less attention, and no x-ray structure has been provided. We have previously co-crystallized PTP1B with a number of low molecular weight inhibitors that inhibit TC-PTP with similar efficiency. Unexpectedly, we were not able to co-crystallize TC-PTP with the same set of inhibitors. This seems to be due to a multimerization process where residues 130-132, the DDQ loop, from one molecule is inserted into the active site of the neighboring molecule, resulting in a continuous string of interacting TC-PTP molecules. Importantly, despite the high degree of functional and structural similarity between TC-PTP and PTP1B, we have been able to identify areas close to the active site that might be addressed to develop selective inhibitors of each enzyme. 相似文献
63.
Pfeiffer A Kunkel T Hiltbrunner A Neuhaus G Wolf I Speth V Adam E Nagy F Schäfer E 《The Plant journal : for cell and molecular biology》2009,57(4):680-689
Translocation from the cytosol to the nucleus is an essential step in phytochrome (phy) signal transduction. In the case of phytochrome A (phyA), this step occurs with the help of FHY1 (far-red-elongated hypocotyl 1), a specific transport protein. To investigate the components involved in phyA transport, we used a cell-free system that facilitates the controlled addition of transport factors. For this purpose, we isolated nuclei from the unicellular green algae Acetabularia acetabulum . These nuclei are up to 100 μm in diameter and allow easy detection of imported proteins. Experiments with isolated nuclei of Acetabularia showed that FHY1 is sufficient for phyA transport. The reconstituted system demonstrates all the characteristics of phytochrome transport in Arabidopsis thaliana . In addition, FHY1 was also actively exported from the nucleus, consistent with its role as a shuttle protein in plants. Therefore, we believe that isolated Acetabularia nuclei may be used as a general tool to study nuclear transport of plant proteins. 相似文献
64.
Susanne Roth Mobarak Abu Mraheil Winfried Barchet Jan Böttcher Torsten Hain Sergej Geiger Yoshihiro Hayakawa Jörg H Fritz Filiz Civril Karl‐Peter Hopfner Christian Kurts Jürgen Ruland Gunther Hartmann Trinad Chakraborty Percy A Knolle 《The EMBO journal》2012,31(21):4153-4164
Immunity against infection with Listeria monocytogenes is not achieved from innate immune stimulation by contact with killed but requires viable Listeria gaining access to the cytosol of infected cells. It has remained ill‐defined how such immune sensing of live Listeria occurs. Here, we report that efficient cytosolic immune sensing requires access of nucleic acids derived from live Listeria to the cytoplasm of infected cells. We found that Listeria released nucleic acids and that such secreted bacterial RNA/DNA was recognized by the cytosolic sensors RIG‐I, MDA5 and STING thereby triggering interferon β production. Secreted Listeria nucleic acids also caused RIG‐I‐dependent IL‐1β‐production and inflammasome activation. The signalling molecule CARD9 contributed to IL‐1β production in response to secreted nucleic acids. In conclusion, cytosolic recognition of secreted bacterial nucleic acids by RIG‐I provides a mechanistic explanation for efficient induction of immunity by live bacteria. 相似文献
65.
An improved approach for transformation of plant cells by microinjection: molecular and genetic analysis 总被引:4,自引:0,他引:4
Martin Schnorf Gabriele Neuhaus-Url Alessandro Galli Shigeru Iida Ingo Potrykus Gunther Neuhaus 《Transgenic research》1991,1(1):23-30
A new culture method for the injection of tobacco mesophyll protoplasts has been established. The protoplasts are embedded
in a thin layer of alginate and are nourished from the medium in the underlying basislayer. In the alginate layer the protoplasts
regenerate to calli at a frequency of up to 80%. Embedded protoplasts can be selected either with 50 mg l−1 kanamycin or 5 mg l−1 paromomycin. Single resistant cells can be recovered from about 10 000 sensitive cells in one alginate layer. Injection of
theneo gene (coding for neomycin phosphotransferase II) into protoplast derived single cells in the alginate layer results in kanamycin
resistant colonies that can be regenerated to mature plants. These plants express the neomycin phosphotransferase as shown
by enzyme activity assay. The integration of the transgene into the plant genome could be proved by Southern hybridization
to high molecular weight DNA. With this culture method 100 cells can be injected per hour. Transformation frequencies range
from 2 to 20%. In crossing experiments, it was shown that the foreign gene is transmitted to the next generation in a Mendelian
fashion. 相似文献
66.
67.
We present a multiview selective-plane illumination microscope (MuVi-SPIM), comprising two detection and illumination objective lenses, that allows rapid in toto fluorescence imaging of biological specimens with subcellular resolution. The fixed geometrical arrangement of the imaging branches enables multiview data fusion in real time. The high speed of MuVi-SPIM allows faithful tracking of nuclei and cell shape changes, which we demonstrate through in toto imaging of the embryonic development of Drosophila melanogaster. 相似文献
68.
Nitrogen‐Doped CNx/CNTs Heteroelectrocatalysts for Highly Efficient Dye‐Sensitized Solar Cells 下载免费PDF全文
Aabhash Shrestha Munkhbayar Batmunkh Cameron J. Shearer Yanting Yin Gunther G. Andersson Joseph G. Shapter Shizhang Qiao Sheng Dai 《Liver Transplantation》2017,7(8)
The use of polydopamine as a nitrogen containing precursor to generate catalytically active nitrogen‐doped carbon (CNx) materials on carbon nanotubes (CNTs) is reported. These N‐doped CNx/CNT materials display excellent electrocatalytic activity toward the reduction of triiodide electrolyte in dye‐sensitized solar cells (DSSCs). Further, the influence of various synthesis parameters on the catalytic performance of CNx/CNTs is investigated in detail. The best performing device fabricated with the CNx/CNTs material delivers power conversion efficiency of 7.3%, which is comparable or slightly higher than that of Pt (7.1%) counter electrode‐based DSSC. These CNx/CNTs materials show great potential to address the issues associated with the Pt electrocatalyst including the high cost and scarcity. 相似文献
69.
70.
Susanne Klumpp Nien Tze MaNicole Bäumer Gunther BechmannJosef Krieglstein 《Biochimica et Biophysica Acta - Proteins and Proteomics》2010,1804(1):206-211
There is increasing evidence that reversible phosphorylation of histidine residues regulates numerous important cellular processes. The first protein histidine phosphatase (PHP) from vertebrates was discovered just recently. Here, we report on amino acids and domains essential for activity of PHP. Point mutations of conserved residues and deletions of the N- and C-termini of PHP were analyzed using [32P-his]ATP-citrate lyase as a substrate. Individual or joint replacement of all cysteine residues by alanine did not affect PHP activity. Deletion of 9 N-terminal amino acids resulted in inactive PHP. Furthermore, only 4 C-terminal residues could be deleted without losing PHP activity. Single or multiple mutations of the glycine-rich domain (Gly75, Gly77) of a putative nucleotide binding site of PHP (GxGxxG/S) caused inactivation of PHP. Wildtype PHP could be labeled with [α-32P]ATP. Such radiolabeling was not detectable for catalytically inactive PHP-G75A and PHP-G77A. These data suggest further studies on the interaction between PHP and ATP. 相似文献