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排序方式: 共有104条查询结果,搜索用时 15 毫秒
1.
Activation of neutrophil calpain following its translocation to the plasma membrane induced by phorbol ester or fMet-Leu-Phe 总被引:5,自引:0,他引:5
S Pontremoli E Melloni F Salamino M Patrone M Michetti B L Horecker 《Biochemical and biophysical research communications》1989,160(2):737-743
Stimulation of human neutrophils with phorbol myristate acetate or fMet-Leu-Phe results in translocation to the plasma membrane of approximately 25-40% of the cellular calpain activity. In the membrane-bound form the Ca2+-requirement for proteolytic activity is substantially reduced. An anti-calpain monoclonal antibody that is internalized by stimulated neutrophils is recovered in the same subcellular fraction that contains the membrane-bound calpain, apparently in the form of pinocytotic vesicles. When both monoclonal antibody and calpain were present in these vesicles, a pronounced inhibition of the membrane bound proteinase activity was observed. These results provide an explanation for the previously observed inhibitory effect of the monoclonal antibody on intracellular calpain activity and on the concomitant inhibition of granule exocytosis. The activated calpain associated with the plasma membrane compartment is therefore identified as the form specifically involved in mediating the physiological responses. 相似文献
2.
The involvement of calpain in the activation of protein kinase C in neutrophils stimulated by phorbol myristic acid 总被引:27,自引:0,他引:27
E Melloni S Pontremoli M Michetti O Sacco B Sparatore B L Horecker 《The Journal of biological chemistry》1986,261(9):4101-4105
The Ca2+/phospholipid-dependent protein kinase (protein kinase C) of human neutrophils is converted to a proteolytically modified Ca2+/phospholipid-independent form (Inoue, M., Kishimoto, A., Takai, Y.U., and Nishizuka, Y. (1977) J. Biol. Chem. 252, 7610-7616) on incubation with neutrophil membranes in the presence of micromolar concentrations of Ca2+ and an endogenous Ca2+-requiring proteinase (Melloni, E., Pontremoli, S., Michetti, M., Sacco, O., Sparatore, B., Salamino, F., and Horecker, B. L. (1985) Proc. Natl. Acad. Sci. U. S. A. 82, 6435-6439). We have now demonstrated the appearance of a similar Ca2+/phospholipid-independent kinase in intact human neutrophils stimulated by phorbol 12-myristate 13-acetate (PMA). The following evidence supports the conclusion that the Ca2+/phospholipid-independent protein kinase recovered from the PMA-treated cells is a proteolytically modified form of the "native" protein kinase C. 1) In cells exposed to PMA, the rate of disappearance of Ca2+/phospholipid-dependent protein kinase C activity is correlated with the rate of appearance of the Ca2+/phospholipid-independent kinase. 2) The chromatographic behavior of the new protein kinase and its molecular size (approximately 65 kDa) are identical to those previously reported for the proteolytically modified form of protein kinase C. 3) The modified protein kinase no longer binds to the cell membrane and is recovered almost entirely in the cytosol fraction. 4) In neutrophils preloaded with inhibitors of the Ca2+-requiring proteinase, stimulation with PMA results in translocation of protein kinase C from the cytosol fraction to the particulate fraction, but the appearance of the soluble, Ca2+/phospholipid-dependent form is prevented. We conclude that binding of protein kinase C to the plasma membrane and its proteolytic conversion are related, but independent, processes both elicited by exposure of neutrophils to the phorbol ester. Proteolytic cleavage of the membrane-bound protein kinase C provides an alternative mechanism for its activation and may account for certain of the cellular responses observed in PMA-stimulated neutrophils. 相似文献
3.
Following association to the membrane, human erythrocyte procalpain is converted and released as fully active calpain 总被引:2,自引:0,他引:2
S Pontremoli F Salamino B Sparatore M Michetti O Sacco E Melloni 《Biochimica et biophysica acta》1985,831(3):335-339
When exposed to inside-out human erythrocyte vesicles, in the presence of micromolar Ca2+, the 80 kDa catalytic subunit of procalpain is processed through three successive and sequential steps. These include binding to the cytosolic surface of the membrane, followed by a very rapid conversion into the 75 kDa active subunit, and ultimately by spontaneous and complete release of this active proteinase form. Binding to the membranes is competitively inhibited by the endogenous natural inhibitor through the formation of the proteinase-inhibitor complex, in which form the 80 kDa subunit can no longer be associated to the membranes. Calcium ions and the natural endogenous inhibitor appear to be crucially involved in the modulation of this novel membrane-bound mediated activation of human red cell procalpain. 相似文献
4.
Binding to erythrocyte membrane is the physiological mechanism for activation of Ca2+-dependent neutral proteinase 总被引:6,自引:0,他引:6
S Pontremoli E Melloni B Sparatore F Salamino M Michetti O Sacco B L Horecker 《Biochemical and biophysical research communications》1985,128(1):331-338
In the presence of micromolar concentrations of Ca2+ the catalytic 80 kDa subunit of human erythrocyte procalpain binds to the cytosolic surface of the erythrocyte membrane. Binding is rapid, highly specific and is reversed by the removal of Ca2+. In the bound form the 80 kDa catalytic subunit undergoes a rapid conversion to calpain, the active 75 kDa Ca2+-requiring proteinase. The activated proteinase produces extensive degradation of membrane components, particularly of band 4.1 and 2.1 proteins. Binding to membranes may represent an obligatory physiological mechanism for the conversion of procalpain to calpain. 相似文献
5.
Ca2+-dependent neutral proteinase from human erythrocytes: activation by Ca2+ ions and substrate and regulation by the endogenous inhibitor 总被引:4,自引:0,他引:4
E Melloni F Salamino B Sparatore M Michetti S Pontremoli 《Biochemistry international》1984,8(4):477-489
Ca2+-dependent neutral proteinase purifies from human erythrocytes as an inactive proenzyme, that can be converted in an active low Ca2+ requiring form either by high concentrations of Ca2+ (0.1-1 mM) in the absence of the substrate, or by low concentrations of Ca2+ (1-5 microM) in the presence of digestible substrates. Activation requires dissociation to constituent inactive proenzyme subunits which are then converted to the active proteinase species still retaining their monomeric structure. The activation process produced by high Ca2+ concentrations is controlled by the endogenous inhibitor which also dissociates into constituent subunits in order to exert its inhibitory effect. An additional regulation of the activated proteinase involves an autoproteolytic process, Ca2+ and substrate dependent, producing enzyme inactivation. 相似文献
6.
Detection by S-100 immunolabelling of interdigitating reticulum cells in human thymomas 总被引:5,自引:0,他引:5
L Lauriola F Michetti V M Stolfi G Tallini D Cocchia 《Virchows Archiv. B, Cell pathology including molecular pathology》1984,45(2):187-195
In the light of recent findings concerning the presence of S-100 antigen in interdigitating reticulum cells (IRC's) in the normal human thymus, we investigated the possible presence of these cells in human thymomas. By the unlabelled antibody PAP method, using an anti-S-100 antiserum, both by light and electron microscopy we were able to demonstrate immunolabelled IRC's in the majority of spindle cell and in some round-oval cell thymomas. Keeping in mind the possible role of IRC's in intrathymic T-cell differentiation, the present findings could be relevant in the better comprehension of this thymic neoplasm. 相似文献
7.
Characterization of three rabbit liver lysosomal proteinases with fructose 1,6-bisphosphatase converting enzyme activity 总被引:3,自引:0,他引:3
E Melloni S Pontremoli F Salamino B Sparatore M Michetti B L Horecker 《Archives of biochemistry and biophysics》1981,208(1):175-183
A large number of nucleoside analogs have been found to inactivate S-adenosylhomocysteine (AdoHcy) hydrolase in a time-dependent irreversible manner. There are two classes of these irreversible inhibitors: (A) analogs that inactivate the enzyme in a pseudofirst-order process and are devoid of any side chain at the 5′-OH group; (B) analogs that inactivate the enzyme in a time-dependent but curvilinear process, and generally have a side chain at the 5′ position. Among the more potent irreversible inhibitors are 2-chloroadenosine, 9-β-d-arabinofuranosyladenine (Ara-A), and (±)aristeromycin. Release of adenine base from adenosine or Ara-A in the presence of AdoHcy hydrolase was observed, thus supporting the proposed catalytic mechanism of AdoHcy hydrolase, that entails the transient formation of 3′-ketoadenosine during enzymatic catalysis of either the formation or hydrolysis of AdoHcy. Both Ara-A and adenosine may exert their irreversible inactivation by a suicide mechanism, but nucleosides such as 5′-iodo-5′-deoxyadenosine and 3′-deoxyadenosine are probably strictly irreversible inhibitors per se in view of the catalytic mechanism proposed for AdoHcy hydrolase. Labeling of AdoHcy hydrolase, perhaps covalent in nature, by radioactive Ara-A and adenosine was demonstrated by gel electrophoresis. 相似文献
8.
The selective vulnerability of specific neuronal subpopulations to trimethyltin (TMT), an organotin compound with neurotoxicant effects selectively involving the limbic system and especially marked in the hippocampus, makes it useful to obtain in vivo models of neurodegeneration associated with behavioural alterations, such as hyperactivity and aggression, cognitive impairment as well as temporal lobe epilepsy. TMT has been widely used to study neuronal and glial factors involved in selective neuronal death, as well as the molecular mechanisms leading to hippocampal neurodegeneration (including neuroinflammation, excitotoxicity, intracellular calcium overload, mitochondrial dysfunction and oxidative stress). It also offers a valuable instrument to study the cell–cell interactions and signalling pathways that modulate injury-induced neurogenesis, including the involvement of newly generated neurons in the possible repair processes. Since TMT appears to be a useful tool to damage the brain and study the various responses to damage, this review summarises current data from in vivo and in vitro studies on neuroprotective strategies to counteract TMT-induced neuronal death, that may be useful to elucidate the role of putative candidates for translational medical research on neurodegenerative diseases. 相似文献
9.
10.
E Melloni B Sparatore F Salamino M Michetti S Pontremoli 《Biochemical and biophysical research communications》1982,107(3):1053-1059
The soluble neutral proteinase of human erythrocytes dissociates into constituent subunits of 80k and 30k in the presence of mM concentrations of Ca2+. Similarly the soluble natural inhibitor of this proteinase, of approximate molecular weight 240k, is dissociated into 60k subunits by mM concentrations of Ca2+. Removal of Ca2+ restores the native oligomeric structure of the proteinase and of the natural inhibitor. The formation of the native active enzyme or of the inactive enzyme-inhibitor complex depends on reversible association-dissociation processes mediated by Ca2+ concentration. 相似文献