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81.
The relationship between the binding patterns of soybean agglutinin, peanut agglutinin (both in their native (unaggregated) form and in their polymerized form), and of Phaseolus vulgaris leucoagglutinin, to neuraminidase-treated lymphocytes from different sources, and the mitogenic activity of these lectins, was studied. In all cases investigated, binding of a lectin to lymphocytes which resulted in stimulation was a positive cooperative process. Our findings support the assumption that clustering of receptors and conformational changes in membrane structure are prerequisites for mitogenic stimulation. 相似文献
82.
Using a substrate-stimulated amino acid efflux system, it has been shown that the “Ly+” and “L” amino acid transport systems of mouse embryo cells in culture are differentially inhibited by parachloromercuribenzene sulfonate (PCMB-S) and the photoaffinity probe 4-fluoro-3-nitrophenylazide (FNPA). Three types of evidence support the conclusion that these transport systems are mediated by separate carrier proteins. (1) The specificity of substrate-stimulated efflux is high for each system; (2) PCMB-S inhibits l-phenylalanine and l-leucine stimulated l-[3H]phenylalanine efflux with no effect on l-lysine stimulated l-[3H]lysine efflux, and (3) the photoaffinity probe FNPA inhibits l-lysine efflux with little effect on the l-phenylalanine-stimulated efflux. 相似文献
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J F Hare 《Biochemical and biophysical research communications》1975,66(4):1329-1337
The energy transducing adenosine 5′-triphosphatase (ATPase) complex was extracted with deoxycholate from membranes and purified 20–25 fold. The detergent-solubilized ATPase complex was inhibited more than 80% by dicyclohexylcarbodiimide (DCCD). Its sedimentation velocity coefficient was 14.7s in the presence of deoxycholate. Phospholipid stimulated its hydrolytic activity and maximized DCCD sensitivity. These parameters clearly differentiate the ATPase complex from the DCCD-insensitive, soluble ATPase prepared by extraction with EDTA at low ionic strength. The purified ATPase complex showed twelve discrete bands on lauryl sulfate gel electrophoresis. Five of these components co-electrophresed with subunits of soluble ATPase. Of the seven additional components, primarily two were precipitated with antibody to soluble ATPase. The protein which specifically reacts with DCCD co-migrated with one of these subunits. 相似文献
85.
The comparison of the proteins of rat liver rough membrane after stripping with EDTA or KCl-puromycin by two dimensional gel electrophoresis is described. By stripping the membrane with EDTA, most of the basic ribosomal proteins are still attached to the membrane; in contrast to the EDTA stripping method, treatment with KCl-puromycin removes most of the ribosomal proteins and does not remove any of the membranal proteins. 相似文献
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Charles A. Berdan Karel A. Erion Nathan E. Burritt Barbara E. Corkey Jude T. Deeney 《PloS one》2016,11(2)
Lipid signals derived from lipolysis and membrane phospholipids play an important role in glucose-stimulated insulin secretion (GSIS), though the exact secondary signals remain unclear. Previous reports have documented a stimulatory role of exogenously added mono-acyl-glycerol (MAG) on insulin secretion from cultured β-cells and islets. In this report we have determined effects of increasing intracellular MAG in the β-cell by inhibiting mono-acyl-glycerol lipase (MGL) activity, which catalyzes the final step in triacylglycerol breakdown, namely the hydrolysis of MAG to glycerol and free fatty acid (FA). To determine the role of MGL in GSIS, we used three different pharmacological agents (JZL184, MJN110 and URB602). All three inhibited GSIS and depolarization-induced insulin secretion in INS-1 (832/13). JZL184 significantly inhibited both GSIS and depolarization-induced insulin secretion in rat islets. JZL184 significantly decreased lipolysis and increased both mono- and diacyglycerol species in INS-1 cells. Analysis of the kinetics of GSIS showed that inhibition was greater during the sustained phase of secretion. A similar pattern was observed in the response of Ca2+ to glucose and depolarization but to a lesser degree suggesting that altered Ca2+ handling alone could not explain the reduction in insulin secretion. In addition, a significant reduction in long chain-CoA (LC-CoA) was observed in INS-1 cells at both basal and stimulatory glucose following inhibition of MGL. Our data implicate an important role for MGL in insulin secretion. 相似文献
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