Abstract A cytotoxin protein was isolated from the sodium dodecyl sulphate (SDS)-solubilized extract of the stable L forms of Salmonella typhimurium by ion-retardation chromatography, ion-exchange chromatography, isoelectric focusing and gel filtration. The purified toxin, with a molecular mass of 32 kDa and with isoelectric point of 6.4, was thermolabile and trypsin-sensitive. Against mouse macrophages, its cytolytic effect was detectable in vitro at concentrations higher than 0.7 μg/ml, with a complete lysis obtained at 5 μg/ml. In contrast, it stimulated C3H/HeJ macrophages in the dose range of 0.1–0.5 μg/ml to allow the cell to respond to endotoxin, resulting in the significant production of tumor necrosis factor α. By Northern blot analysis, this effect was detectable at a dose as low as 0.01 μg/ml. These findings suggest that the transformation of bacillary S. typhimurium into L forms in vivo may induce alterations in host resistance against murine typhoid. 相似文献
Stress fibers (SFs) in cells transmit external forces to cell nuclei, altering the DNA structure, gene expression, and cell activity. To determine whether SFs are involved in mechanosignal transduction upon intraluminal pressure, this study investigated the SF direction in smooth muscle cells (SMCs) in aortic tissue and strain in the SF direction. Aortic tissues were fixed under physiological pressure of 120 mmHg. First, we observed fluorescently labeled SFs using two-photon microscopy. It was revealed that SFs in the same smooth muscle layers were aligned in almost the same direction, and the absolute value of the alignment angle from the circumferential direction was 16.8° ± 5.2° (n = 96, mean ± SD). Second, we quantified the strain field in the aortic tissue in reference to photo-bleached markers. It was found in the radial-circumferential plane that the largest strain direction was − 21.3° ± 11.1°, and the zero normal strain direction was 28.1° ± 10.2°. Thus, the SFs in aortic SMCs were not in line with neither the largest strain direction nor the zero strain direction, although their orientation was relatively close to the zero strain direction. These results suggest that SFs in aortic SMCs undergo stretch, but not maximal and transmit the force to nuclei under intraluminal pressure.
The main characteristic changes observed in Alzheimer's disease (AD) are the presence of neurofibrillary tangles and the deposition of amyloid A4 peptides. The most abundant amyloid A4 peptide species in AD (which we tentatively named A4') is composed of 39 amino acids, which is devoid of the 3 N-terminal amino acids, Asp-Ala-Glu, of the originally reported A4 peptide. We synthesized a model peptide substrate, Suc-Ala-Glu-methylcoumarinamide (MCA), to identify the proteinase that splits the A4' peptide. DEAE-cellulose column chromatography of rat liver and porcine brain extracts showed that only one peak material digested the synthetic substrate at pH 8. The results for the final preparation indicate that the Suc-Ala-Glu-MCA-degrading enzyme is a high-molecular-mass proteinase, with a molecular mass of above 500,000, and is composed of several low-molecular-mass subunits. These results suggest that a non-lysosomal multicatalytic proteinase (we named this enzyme ingensin (ingens = large in Latin). Ishiura, S. et al. (1985) FEBS Lett. 189, 119-123) catalyzes the above reaction. Antiserum against the purified multicatalytic proteinase, ingensin, crossreacted with the purified Suc-Ala-Glu-MCA-degrading proteinase. It is likely that ingensin shows a similar action toward amyloid precursor protein (APP) in vivo. 相似文献
In a previous study we isolated homologues of new oleanane-type triterpene
glycosides from leaves of Staurogyne merguensis Kuntze and named them
strogins. Strogins themselves have a sweet taste (sweet activity), which
diminishes in a few minutes. Subsequent application of cold water to the
mouth then elicits a sweet taste (sweetness-inducing activity). In the
present study we systematically examined the properties of the sweet and
sweetness-inducing activities of strogins. Strogins 1, 2 and 4 had both the
sweet and sweetness-inducing activities, while strogins 3 and 5 had no
activities. The sweetness- inducing activity in response to cold water
lasted for 1 h for strogin 2 and 2 h for strogins 1 and 4. The
sweetness-inducing activity was immediately diminished by application of
gamma-cyclodextrin to the mouth after strogins were held in the mouth. It
seems that the strogins were adsorbed on the gustatory receptor membranes
and eliminated by inclusion activity of gamma-cyclodextrin. The structure
of strogin resembles that of gymnemic acid, which has antisweet activity.
There was competition between strogin 1 and gymnemic acid; treatment of the
tongue with strogin 1 before application of Gymnema extract to the mouth
reduced the antisweet activity. While the sweetness-inducing activity of
curculin in response to water was suppressed by the presence of divalent
cations such as Ca2+ or Mg2+, that of strogin was not suppressed by the
divalent cations. The changes in the inactive complex between strogin and
the sweet receptor site in the adaptation state into the active complex
induced by cold stimulation were discussed.
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The multicatalytic proteinase, ingensin, was purified to homogeneity from chicken liver. rRNA-degrading activity was co-eluted with the purified multicatalytic proteinase from a TSK-3000SW column. This RNA-degrading activity was inactivated by heat treatment and the addition of a low concentration of SDS. Therefore, the RNA-degrading activity co-eluted with the multicatalytic proteinase was not due to contamination by low-molecular-mass RNases. These results strongly suggest that this RNA-degrading activity was tightly associated with the multicatalytic proteinase, ingensin. 相似文献