Active site phosphoryl groups in the biphosphorylated phosphotransferase complex reveal dynamics in a millisecond time scale

AbstractThe N-terminal domain of Enzyme I (EIN) and phosphocarrier HPr can form a biphosphorylated complex when they are both phosphorylated by excess cellular phosphoenolpyruvate. Here we show that the electrostatic repulsion between the phosphoryl groups in the biphosphorylated complex results in characteristic dynamics at the active site in a millisecond time scale. The dynamics is localized to phospho-His15 and the stabilizing backbone amide groups of HPr, and does not impact on the phospho-His189 of EIN. The dynamics occurs with the kex of ∼500s−1 which compares to the phosphoryl transfer rate of ∼850s−1 between EIN and HPr. The conformational dynamics in HPr may be important for its phosphotransfer reactions with multiple partner proteins.Structured summary of protein interactionsEIN and HPr bind by nuclear magnetic resonance (View Interaction)

Lateral resistance of mass timber shear wall connected by withdrawal-type connectors

In this study, the lateral resistances of mass timber shear walls were investigated for seismic design. The lateral resistances were predicted by kinematic models with mechanical properties of connectors, and compared with experimental data. Four out of 7 shear wall specimens consisted of a single Ply-lam panel and withdrawal-type connectors. Three out of 7 shear wall specimens consisted of two panels made by dividing a single panel in half. The divided panels were connected by 2 or 4 connectors like a single panel before being divided. The applied vertical load was 0, 24, or 120 kN, and the number of connectors for connecting the Ply-lam wall-to-floor was 2 or 4. As a result, the tested data were 6.3 to 52.7% higher than the predicted value by kinematic models, and it means that the lateral resistance can be designed by the behavior of the connector, and the prediction will be safe. The effects of wall-to-wall connectors, wall-to-floor connectors and vertical loads on the shear wall were analyzed with the experimental data.This project was funded by the Korea Forest Service (Korea Forestry Promotion Institute) and the Ministry of Land, Infrastructure and Transport (the Korea Agency for Infrastructure Technology Advancement)

Sensitivity of censored data analysis to determine the characteristic value of structural timber

In structural timber tests, unintended failure mechanisms occur frequently in specimens and their results are called censored data. There are two censored data analysis: censored maximum likelihood estimation (CMLE) and Kaplan–Meier (KM) method. In this study, the precision of the censored data analysis was investigated to determine the characteristic value, 5th percentile value, of the structural timber. The results show that (1) the 5th percentile value was underestimated by ordinary data analysis methods; maximum likelihood estimation (MLE) and Order statistics. (2) CMLE with 30% lower tail censored data and KM method provided much more precise 5th percentile value. (3) The amount of under-measurement (5 MPa, 10 MPa, and 15 MPa in this simulation study) did not show significant effect on the 5th percentile determination in CMLE and KM method, but the proportion of censored data (percentage of unintended failure specimen; 10%, 20%, 30%, and 40%) affected the determination of 5th percentile value. (4) CMLE with 30% lower tail censored data and KM method showed good agreement in case that the data included unintended failure data up to 20%.(1) This work was supported by Research Resettlement Fund for the new fac‑ulty of Seoul National University. (2) This work was supported by the National Research Foundation of Korea (NRF) grant funded by the Korea government (MSIT) (No. 2017R1A2B1010748)

Prediction of withdrawal resistance for a screw in hybrid cross-laminated timber

The aim of this study was to predict the withdrawal resistance of a screw in hybrid cross-laminated timber (CLT) composed of two types of lamina layers. A theoretical model to predict the withdrawal resistance was developed from the shear mechanism between a screw and the layers in hybrid CLT. The parameters for the developed model were the withdrawal stiffness and strength that occurs when a screw is withdrawn, and the penetration depth of a screw in layers of a wood material. The prediction model was validated with an experimental test. Screws with two different diameters and lengths (Ø6.5 × 65mm and Ø8.0 × 100mm) were inserted in a panel composed of solid wood and plywood layers, and the withdrawal resistances of the screws were evaluated. At least 30 specimens for each group were tested to derive the lower 5th percentile values. As a result, the developed model predictions were 86–88% of the lower 5th percentile values of hybrid CLT from the properties of the lamina layer. This shows that the withdrawal resistance of hybrid CLT can be designed from the properties of its layer.This project was funded by the Korea Forest Service (Korea Forestry Promotion Institute) and the Ministry of Land, Infrastructure and Transport (the Korea Agency for Infrastructure Technology Advancement)

Solubility enhancement of aggregation-prone heterologous proteins by fusion expression using stress-responsive Escherichia coli protein, RpoS

<p>Abstract</p> <p>Background</p> <p>The most efficient method for enhancing solubility of recombinant proteins appears to use the fusion expression partners. Although commercial fusion partners including maltose binding protein and glutathione-<it>S</it>-transferase have shown good performance in enhancing the solubility, they cannot be used for the proprietory production of commercially value-added proteins and likely cannot serve as universal helpers to solve all protein solubility and folding issues. Thus, novel fusion partners will continue to be developed through systematic investigations including proteome mining presented in this study.</p> <p>Results</p> <p>We analyzed the <it>Escherichia coli </it>proteome response to the exogenous stress of guanidine hydrochloride using 2-dimensional gel electrophoresis and found that RpoS (RNA polymerase sigma factor) was significantly stress responsive. While under the stress condition the total number of soluble proteins decreased by about 7 %, but a 6-fold increase in the level of RpoS was observed, indicating that RpoS is a stress-induced protein. As an N-terminus fusion expression partner, RpoS increased significantly the solubility of many aggregation-prone heterologous proteins in <it>E. coli </it>cytoplasm, indicating that RpoS is a very effective solubility enhancer for the synthesis of many recombinant proteins. RpoS was also well suited for the production of a biologically active fusion mutant of <it>Pseudomonas putida </it>cutinase.</p> <p>Conclusion</p> <p>RpoS is highly effective as a strong solubility enhancer for aggregation-prone heterologous proteins when it is used as a fusion expression partner in an <it>E. coli </it>expression system. The results of these findings may, therefore, be useful in the production of other biologically active industrial enzymes, as successfully demonstrated by cutinase.</p

Optical repumping of triplet PP-states enhances magneto-optical trapping of ytterbium atoms

Radiative decay from the excited $^1P_1$ state to metastable $^3P_2$ and $^3P_0$ states is expected to limit attainable trapped atomic population in a magneto-optic trap of ytterbium (Yb) atoms. In experiments we have carried out with optical repumping of $^3P_{0,2}$ states to $^3P_1$, we observe enhancement of trapped atoms yield in the excited $^1P_1$ state. The individual decay rate to each metastable state is measured and the results show an excellent agreement with the theoretical values.Comment: 5 pages, 5 figure

Effective Action and Schwinger Pair Production in Scalar QED

Some astrophysical objects are supposed to have very strong electromagnetic fields above the critical strength. Quantum fluctuations due to strong electromagnetic fields modify the Maxwell theory and particularly electric fields make the vacuum unstable against pair production of charged particles. We study the strong field effect such as the effective action and the Schwinger pair production in scalar QED.Comment: RevTex 6 pages, no figure; Proceedings of APCTP Winter School on Black Hole Astrophysics 2008, Jan 24-29, 200

A two-photon fluorescent probe for lysosomal zinc ions

The selective detection of zinc ions in lysosomes over that in cytosol is achieved with a fluorescent probe, which enabled the fluorescence imaging of endogenous zinc ions in lysosomes of NIH 3T3 cells as well as mouse hippocampal tissues by two-photon microscopy under excitation at 900 nm.open

Crystallization and preliminary X-ray crystallographic studies of the ice-binding protein from the Antarctic yeast Leucosporidium sp. AY30. Corrigendum

A correction to the article by Park et al. [(2011). Acta Cryst. F67, 800–802]

Proximal Subungual Onychomycosis in a Patient with Classic Kaposi Sarcoma Caused by Trichophyton rubrum

A 58-year-old man presented with whitish patches on both great toenails for four weeks prior to visiting our hospital; the patches spread rapidly to other finger- and toe-nails. Prior to presentation, the patient had been diagnosed with idiopathic thrombocytopenic purpura two months ago and Kaposi's sarcoma three weeks ago. The patient was treated with human immunoglobulin for five days, and then received prednisolone 40 mg bid. Serology showed that the patient was negative for HIV and results of other laboratory tests were normal. The KOH slide preparation of the nail scraping showed long septated hyphae and numerous arthrospores. The fungus culture revealed whitish downy colonies on the front side and wine-red reverse pigmentation on Sabouraud's dextrose agar. Trichophyton rubrum was isolated on fungus culture and slide culture. The internal transcribed space (ITS) regions of ribosomal DNA of the cultured fungus were identical to Trichophyton rubrum. Proximal subungual onychomycosis (PSO) is the rarest form of onychomycosis. PSO initially presents as whitish patch(es) on the proximal side of the nail plate(s). Because PSO shows whitish to yellowish patches on the nail plate, the result of KOH examination of nail scrapings from the nail plate is almost always negative. Herein, we report on a case of multiple PSO in a patient with classic Kaposi sarcoma and suggest a method for easy KOH scraping on PSO