396 research outputs found
Molecular Electroporation and the Transduction of Oligoarginines
Certain short polycations, such as TAT and polyarginine, rapidly pass through
the plasma membranes of mammalian cells by an unknown mechanism called
transduction as well as by endocytosis and macropinocytosis. These
cell-penetrating peptides (CPPs) promise to be medically useful when fused to
biologically active peptides. I offer a simple model in which one or more CPPs
and the phosphatidylserines of the inner leaflet form a kind of capacitor with
a voltage in excess of 180 mV, high enough to create a molecular electropore.
The model is consistent with an empirical upper limit on the cargo peptide of
40--60 amino acids and with experimental data on how the transduction of a
polyarginine-fluorophore into mouse C2C12 myoblasts depends on the number of
arginines in the CPP and on the CPP concentration. The model makes three
testable predictions.Comment: 15 pages, 5 figure
Simple Model of the Transduction of Cell-Penetrating Peptides
Cell-penetrating peptides (CPPs) such as HIV's trans-activating
transcriptional activator (TAT) and polyarginine rapidly pass through the
plasma membranes of mammalian cells by an unknown mechanism called
transduction. They may be medically useful when fused to well-chosen chains of
fewer than about 35 amino acids. I offer a simple model of transduction in
which phosphatidylserines and CPPs effectively form two plates of a capacitor
with a voltage sufficient to cause the formation of transient pores
(electroporation). The model is consistent with experimental data on the
transduction of oligoarginine into mouse C2-C12 myoblasts and makes three
testable predictions.Comment: Seven pages. For a more complete version including the effects of
counterions, see arXiv:0810.2358v3 [q-bio.BM
Validation of the High Performance Conduction-Cooled Prototype LTS Pulse Coil for UPS-SMES
A conduction-cooled low temperature superconducting (LTS) pulse coil has been developed as a key technology for UPS-SMES. We have been developing a 1 MW, 1 s UPS-SMES for a protection from a momentary voltage drop and an instant power failure. A conduction-cooled LTS pulse coil has excellent characteristics, which are adequate for a short-time uninterruptible power supply (UPS). The LTS coil has better cost performance over the HTS coil at present and the conduction cooling has higher reliability and easier operation than the conventional cooling schemes such as pool boiling with liquid helium or forced flow of supercritical helium. To demonstrate the high performances of the LTS pulse coil, we have fabricated a prototype coil with stored energy of 100 kJ and have conducted cooling and excitation tests. The successful performance test results including current shut-off test with a time constant of 1.3 s and repeated excitation of a triangular waveform with high ramp rate are reporte
Impact of the Wall Conditioning Program on Plasma Performance in NSTX
High performance operating regimes have been achieved on NSTX (National Spherical Torus Experiment) through impurity control and wall-conditioning techniques. These techniques include HeGDC-aided boronization using deuterated trimethylboron, inter-discharge HeGDC, 350 C PFC bake-out followed by D2 and HeGDC, and experiments to test fueling discharges with either a He-trimethylboron mixture or pure trimethylboron. The impact of this impurity and density control program on recent advances in NSTX plasma performance is discussed
Vibrio cholerae Proteome-Wide Screen for Immunostimulatory Proteins Identifies Phosphatidylserine Decarboxylase as a Novel Toll-Like Receptor 4 Agonist
Recognition of conserved bacterial components provides immediate and efficient immune responses and plays a critical role in triggering antigen-specific adaptive immunity. To date, most microbial components that are detected by host innate immune system are non-proteinaceous structural components. In order to identify novel bacterial immunostimulatory proteins, we developed a new high-throughput approach called βEPSIAβ, Expressed Protein Screen for Immune Activators. Out of 3,882 Vibrio cholerae proteins, we identified phosphatidylserine decarboxylase (PSD) as a conserved bacterial protein capable of activating host innate immunity. PSD in concentrations as low as 100 ng/ml stimulated RAW264.7 murine macrophage cells and primary peritoneal macrophage cells to secrete TNFΞ± and IL-6, respectively. PSD-induced proinflammatory response was dependent on the presence of MyD88, a known adaptor molecule for innate immune response. An enzymatically inactive PSD mutant and heat-inactivated PSD induced βΌ40% and βΌ15% of IL-6 production compared to that by native PSD, respectively. This suggests that PSD induces the production of IL-6, in part, via its enzymatic activity. Subsequent receptor screening determined TLR4 as a receptor mediating the PSD-induced proinflammatory response. Moreover, no detectable IL-6 was produced in TLR4-deficient mouse macrophages by PSD. PSD also exhibited a strong adjuvant activity against a co-administered antigen, BSA. Anti-BSA response was decreased in TLR4-deficient mice immunized with BSA in combination with PSD, further proving the role of TLR4 in PSD signaling in vivo. Taken together, these results provide evidence for the identification of V. cholerae PSD as a novel TLR4 agonist and further demonstrate the potential application of PSD as a vaccine adjuvant
Sec16 Defines Endoplasmic Reticulum Exit Sites and is Required for Secretory Cargo Export in Mammalian Cells
The selective export of proteins and lipids from the endoplasmic reticulum (ER) is mediated by the coat protein complex II (COPII) that assembles onto the ER membrane. In higher eukaryotes, COPII proteins assemble at discrete sites on the membrane known as ER exit sites (ERES). Here, we identify Sec16 as the protein that defines ERES in mammalian cells. Sec16 localizes to ERES independent of Sec23/24 and Sec13/31. Overexpression, and to a lesser extent, small interfering RNA depletion of Sec16, both inhibit ER-to-Golgi transport suggesting that Sec16 is required in stoichiometric amounts. Sar1 activity is required to maintain the localization of Sec16 at discrete locations on the ER membrane, probably through preventing its dissociation. Our data suggest that Sar1-GTP-dependent assembly of Sec16 on the ER membrane forms an organized scaffold defining an ERES
A novel synthesis and detection method for cap-associated adenosine modifications in mouse mRNA
A method is described for the detection of certain nucleotide modifications adjacent to the 5' 7-methyl guanosine cap of mRNAs from individual genes. The method quantitatively measures the relative abundance of 2'-O-methyl and N6,2'-O-dimethyladenosine, two of the most common modifications. In order to identify and quantitatify the amounts of N6,2'-O-dimethyladenosine, a novel method for the synthesis of modified adenosine phosphoramidites was developed. This method is a one step synthesis and the product can directly be used for the production of N6,2'-O-dimethyladenosine containing RNA oligonucleotides. The nature of the cap-adjacent nucleotides were shown to be characteristic for mRNAs from individual genes transcribed in liver and testis
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