Katsuhiko Shimada

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We have established a fluorescence method to detect calcium-binding proteins by making use of the quinoline Ca indicator quin2. Authentic calcium-binding proteins were subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis and then electrophoretically transferred onto polyvinylidene difluoride membranes. Transfers were incubated with(More)
Neuroglycan C (NGC), a brain-specific transmembrane proteoglycan, is thought to bear not only chondroitin sulfate but also N- and O-linked oligosaccharides on its core protein. In this study, we isolated and purified NGC from rat brains at various developmental stages by immunoaffinity column chromatography or by immunoprecipitation, and examined the(More)
OBJECTIVE To determine the signal transduction pathways in CD14+ synovial cells from patients with rheumatoid arthritis (RA) after CD40 ligation, and to examine their role in amplifying synovial inflammation in affected joints. METHODS Expression of messenger RNA was analyzed using quantitative reverse transcription-polymerase chain reaction. Cytokines(More)
The first study by nmr of the integral membrane protein, the bacterial light-harvesting (LH) antenna protein LH1 beta, is reported. The photosynthetic apparatus of purple bacteria contains two different kinds of antenna complexes (LH1 and LH2), which consist of two small integral membrane proteins alpha and beta, each of approximately 6 kDa, and(More)
We examined the capacity of tropomyosin molecules regulating the sliding movement of actin filaments on myosin molecules in the presence of ATP molecules to be hydrolyzed. For this objective, we prepared tropomyosin molecules modified to be a little bit stiffer compared to the intact ones by applying a fixed cross-linker between a pair of twisted(More)
Phosphate buffer suspensions of resting Escherichia coli B cells at pH 7.0 were anaerobically exposed to alternating current (a.c.) of 50 Hz at a current density of 600 +/- 60 mA/cm2 and 34 degrees +/- 3 degrees C. The minimum inhibitory concentrations of eight basic dyes: crystal violet, malachite green, brilliant green, fuchsin, methylene blue, toluidine(More)
A system to use fluorometry for the search of protein crystallization buffers was developed. The screening of candidates was done with a use of short gel-filtration column (Sephacryl S-100 HR) linked to the fluorometer. Protein elution was monitored by intrinsic fluorescence or emission from its labels. This method was applied to actin and actin complexes.(More)
We examined both longitudinal and transversal fluctuations of displacements of an actin filament sliding upon Chara myosin molecules. Although the magnitude of transversal fluctuations remained rather independent of ATP concentration, the longitudinal ones were found to increase their magnitude as the concentration increased. In addition, the longitudinal(More)
To elucidate the nature of myospherulosis, the authors tried to produce this state in vitro by mixing vitamin E, oleic acid, linoleic acid, and lanolin with human blood components, such as whole blood, washed erythrocytes, plasma, and fixed erythrocytes, respectively. Myospherulosis was produced in all mixtures in these experiments. The authors concluded(More)
The sliding velocity of actin filaments was found to increase in the presence of ATP analogues. At 0.5 mM ATP, the presence of 2.0 mM of AMP-PNP enhanced the filament velocity from 3.2 up to 4.5 microm/s. However, 2 mM ADP decreased the velocity down to 1.1 microm/s. The results suggest that the complex conformations of myosin cross-bridges interacting with(More)