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In the absence of MgATP, the catalytic subunit of cAMP-dependent protein kinase is irreversibly inhibited by the hydrophobic carbodiimide dicyclohexylcarbodiimide, and this inhibition is most likely due to the formation of a cross-link between a carboxyl group and a lysine residue in the active site (Toner-Webb & Taylor, 1987). In order to identify these cross-linked residues, the catalytic subunit was modified by dicyclohexylcarbodiimide and then treated with acetic anhydride and digested with trypsin. The resulting peptides were resolved by high-performance liquid chromatography. One major absorbing tryptic peptide and one smaller peptide consistently and reproducibly showed a decrease in absorbance after the catalytic subunit had been treated with DCCD. These peptides correspond to residues 166-190 and 57-93, respectively. A unique peptide was isolated from the modified catalytic subunit, and the sequence of this peptide established that the cross-linking occurred between Asp-184 and Lys-72. The cross-linking of these two residues, which were both identified previously as essential residues, confirms the likelihood that each plays a role in the functioning of this enzyme. The fact that Asp-184 and Lys-72 appear to be invariant in all protein kinases further supports the hypothesis that these two residues, located close to one another at the active site of the enzyme, play essential roles in catalysis. 相似文献
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Calculations are presented of the induced electric fields and current densities in the cartilage of the knee produced by a coil applicator developed for applying pulsed magnetic fields to osteoarthritic knees. This applicator produces a sawtooth-like magnetic field waveform composed of a series of 260-micros pulses with a peak to peak magnitude of approximately 0.12 mT in the cartilage region. The simulations were performed using a recently developed 3 dimensional finite difference frequency domain technique for solving Maxwell's equations with an equivalent circuit model. The tissue model was obtained from the anatomically segmented human body model of Gandhi. The temporal peak electric field magnitude was found to be -153 mV/m, averaged within the medial cartilage of the knee for the typical dB/dt excitation levels of this coil. The technique can be extended to analyze other excitation waveforms and applicator designs. 相似文献
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Sigruener A Buechler C Bared SM Grandl M Aslanidis C Ugocsai P Gehrmann M Schmitz G 《Biochemical and biophysical research communications》2007,359(3):723-728
Uptake of modified lipoproteins by macrophages causes foam cell formation and promotes atherosclerosis. Atherogenic lipoproteins are cytotoxic and induce cell death under certain conditions but may also enhance macrophage survival. Macrophages treated with enzymatically modified LDL (E-LDL) were subjected to GeneChip analysis and the antiapoptotic gene TOSO was found induced. TOSO mRNA is upregulated and apoptosis is reduced in E-LDL but not in oxidized LDL (Ox-LDL) loaded macrophages. FLIP(L) abundance was suggested to mediate the antiapoptotic properties of TOSO; however, FLIP(L) was not changed. Ox-LDL is internalized predominantly by scavenger receptors such as CD36 while E-LDL particles are preferentially internalized by Fc- and complement-receptor dependent phagocytosis and internalization of phagobeads by macrophages upregulates TOSO. In COS-7 cells however, phagocytotic activity was not affected by TOSO. These data indicate that E-LDL-generated foam cells are protected from cell death most likely through the expression of TOSO by a FLIP(L) independent mechanism. 相似文献
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Phosphorus regeneration in fresh-water paramecia 总被引:2,自引:0,他引:2
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Identification of aminopyrimidine‐sulfonamides as potent modulators of Wag31‐mediated cell elongation in mycobacteria 下载免费PDF全文
János Pató Gaëlle S. Kolly Jana Korduláková Martin Forbak Joanna C. Evans Rita Székely Jan Rybniker Zuzana Palčeková Júlia Zemanová Isabella Santi François Signorino‐Gelo Liliana Rodrigues Anthony Vocat Adrian S. Covarrubias Monica G. Rengifo Kai Johnsson Sherry Mowbray Joseph Buechler Vincent Delorme Priscille Brodin Graham W. Knott José A. Aínsa Digby F. Warner Katarína Mikušová John D. McKinney Ruben C. Hartkoorn 《Molecular microbiology》2017,103(1):13-25
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Wurm S Neumeier M Weigert J Wanninger J Gerl M Gindner A Schäffler A Aslanidis C Schölmerich J Buechler C 《Cytokine》2008,44(1):185-190
Oral glucose uptake alters the function of immune cells and an elevation of systemic CXCL8 was described. Monocytes secrete high amounts of CXCL8 and therefore it was analyzed whether glucose or insulin upregulate monocytic CXCL8 release. Incubation of monocytes with insulin for 2h induced CXCL8 mRNA and secretion whereas glucose had no effect. Inhibition of the phosphatidylinositol 3-kinase by wortmannin or the mammalian target of rapamycin by rapamycin did not influence insulin-mediated CXCL8 induction. In contrast, blockage of the ERK-specific MAP kinase MEK with PD98059, that prevents phosphorylation of ERK1/ERK2, abrogated insulin-induced CXCL8 release in primary monocytes. To investigate the in vivo effect of oral glucose uptake, monocytes of healthy probands were isolated in the fasted state and 2h after glucose ingestion and CXCL8 mRNA and protein were increased in the latter. CXCL8 was also higher when determined in the cell lysate of leukocytes 2h after glucose uptake whereas plasma CXCL8 levels were significantly reduced. In summary, these data indicate that oral glucose uptake in insulin-sensitive adults is associated with elevated monocytic and reduced systemic CXCL8. 相似文献