共查询到20条相似文献,搜索用时 31 毫秒
1.
Misao Owada Elizabeth F. Neufeld 《Biochemical and biophysical research communications》1982,105(3):814-820
We have examined frozen liver tissue for N-acetylglucosamine-l-phosphotransferase, an enzyme required for the formation of the mannose 6-phosphate recognition marker of lysosomal enzymes. Using [β32P]-UDPGlcNAc and placental β-hexosaminidase B as N-acetylglucosamine l-phosphate donor and acceptor, respectively, we were unable to find activity of the transferase in 100,000 × g membranes prepared from livers of patients with I-cell disease, whereas activity was readily observed in membranes from control livers stored under the same conditions. Yet the activity of several lysosomal enzymes (β-N-acetylglucosaminidase, β-glucuronidase, α-mannosidase and α--iduronidase) was comparable in liver tissue of I-cell patients and controls, and only β-galactosidase activity showed a marked reduction. These results suggest that in contrast to cultured skin fibroblasts, liver may be able to introduce into lysosomes acid hydrolases that lack the mannose 6-phosphate recognition marker. 相似文献
2.
P Dell'Antone 《Biochemical and biophysical research communications》1979,86(1):180-189
When rat liver lysosomes are suspended in a medium containing acridine orange at neutral pH, accumulation of the dye may be observed within the vesicles. The uptake appears driven by a pH gradient between the external medium and the interior of the lysosomes since it is inhibited by NH4+, nigericin and other electroneutral proton-cation exchangers. FCCP is ineffective in inhibiting the uptake. In the presence of Mg++ and anions such as Cl?, ATP promoted a further and more extensive but slower oligomycin and ouabain-insensitive dye uptake, which was also inhibited by FCCP. Very similar results were obtained with neutral red and atebrin. When the rate of the ATP-induced acridine uptake in preparations of different purification grade was compared, it was observed that the uptake rate increased in parallel with lysosomal enzymatic activity. These results suggest that an electrogenic ATP-driven-Mg++ dependent “proton pump” is operating in the lysosomal membrane, as previously proposed. 相似文献
3.
Ten steroids have been compared for their ability to modify the rate of uptake of acridine orange by rat liver and by rat liver lysosomes in vivo. The short-term effects of the ten steroids on the specific activity of a lysosomal enzyme, beta-N-acetylglucosaminidase, were also compared. Five of the ten steroids were administered as tritium-labelled compounds and the concentration of steroids or metabolites was measured in rat liver and liver lysosomes at 2.5h and 3.75h after administration. Cortisone acetate, etiocholanolone (5-beta-androstan-3-alpha-01-17-one) and testosterone accelerate and increase the uptake of acridine orange by rat liver lysosomes. Deoxycorticosterone, corticosterone, triamcinolone (9-alpha-fluoro-11-beta, 17, 21-trihydroxy-16-alpha-methyl-pregna-1, 4-diene-3, 20-dione), estradiol-17-beta and progesterone appear to inhibit the uptake of acridine orange by rat liver lysosomes at 2.5 hours. Cortisol and dexamethasone (9-alpha-fluoro-11-beta, 17, 21-trihydroxy-16-alpha-methyl-pregna-1, 4-diene-3, 20-dione) had little effect. All steroids with the exception of etiocholanolone and deoxycorticosterone increase with the specific activity of beta-N-acetylglucosaminidase in the lysosomal fraction at 2.5h. None of the effects at 2.5h are due to lowered protein levels. Lysosomal concentrations of radioactivity following the administration of tritiated steroids were greated for the glucocorticoids, corticosterone and cortisol. Estradiol-17-beta, progesterone and testosterone showed much lower concentrations of radioactivity in isolated lysosomes. Most of the lysosomal radioactivity (73-96%) was associated with the soluble fraction of the disrupted lysosomes. 相似文献
4.
A membranous ATPase unique to lysosomes 总被引:1,自引:0,他引:1
D L Schneider 《Biochemical and biophysical research communications》1974,61(3):882-888
Intracellular protein degradation presumably occurs in lysosomes and is known to require energy. ATP effects on proteolysis in lysosomes have been demonstrated and were attributed to transport of protons (1) or transport of proteins (2). Accordingly a lysosomal ATPase was anticipated and is reported herein. The membranous nature of the ATPase as well as its specific activity would seemingly warrant its consideration as the direct link between energy and protein catabolism. However, lysosomes display very little ATPase activity and there is no stimulation by uncoupler. Therefore the existence of an ATP-driven proton pump in lysosomes is improbable. 相似文献
5.
Intracellular origin of hormone-sensitive lipolysis in the rat 总被引:2,自引:0,他引:2
The hormone-stimulated hydrolysis of endogenous triglycerides in heart and adipose tissue was found to be inhibited by chloroquine, which is known to accumulate in lysosomes and to inhibit the lysosomal degradation of protein and cholesterolesters.When the triglyceride depôt in heart cells was increased by feeding rats a diet enriched in erucic acid for three days prior to perfusion of the heart, the spontaneous and the norepinephrine stimulated rates of lipolysis were both found to be increased. Both were inhibited by chloroquine. Analysis of trioleoylglycerol hydrolysis in heart homogenates after heparin perfusion revealed the virtual absence of neutral lipase in contrast to an acid lipase activity. The results of this study suggest that lipolytic activity of lysosomal origin is the main source of hormone-sensitive endogenous triacylglycerol hydrolysis. 相似文献
6.
R.W.Scot Skinner Rodney V. Pozderac Raymond E. Counsell Chen-Fu Hsu Paul A. Weinhold 《Steroids》1977,30(1):15-23
2-selena-A-nor-5α-androstan-17β-ol was studied and in the rat prostate gland. The data demonstrates the ability of this compound to selectively complex with the specific receptors of 5α-dihydrotestosfcerone (5α-DHT) in the cytosol and to be retained in the nuclei in an unaltered form. Studies with selenium-75 labeled material suggests that the uptake and localization is similar to endogenous 5α-dihydrotestosterone. 相似文献
7.
Nathan N. Aronson 《Life sciences》1980,27(2):95-104
Recent experimental evidence has been obtained, principally in the laboratory of Glenn Mortimore, that hepatic lysosomes can act as a pool of amino acids during fasting. This pool is generated through , whereby intracellular proteins are somehow captured by the lysosomes and then rapidly hydrolyzed to free amino acids by the lysosomal proteinases. Two important metabolic fates of these lysosomal digestive products can be: 1) conversion of the glucogenic amino acids into glucose, and 2) conversion of trimethyl-lysine into carnitine. The latter metabolite is required to transfer fatty acids to the mitochondrial site of β-oxidation. Most interesting is the observation that glucagon appears to induce lysosomal autophagy and the resulting degradation of intracellular proteins by decreasing the size of amino acid pools in the perfused liver. This effect of the hormone may be directed at the single amino acid glutamine, since adding it alone to the perfusate can prevent the increase in autophagy caused by glucagon. Insulin also rapidly inactivates hepatic autophagy and its ensuing proteolysis. The for the rate of los of autophagic vocuoles from the insulin-treated liver (or animal) is approximately 8 min. Thus, glucagon and insulin actively control intracellular protein catabolism that takes place within hepatic lysosomes, and this regulation by the two hormones may be one of their major molecular effects on gluconegenesis in the liver. 相似文献
8.
Previous reports indicate that alterations of activity of cholinergic neurons are followed by parallel changes in sodium-dependent high affinity choline uptake . These results are consistent with the proposal that this portion of choline uptake is regulatory in the synthesis of ACh. These results also suggest the possibility of utilizing sodium-dependent high affinity choline uptake as a measure of the relative state of cholinergic activity . In this study, we administer a number of drugs reported to alter turnover and release of ACh (both are measures of cholinergic activity , and subsequently examine sodium-dependent high affinity choline uptake . Administration of pentobarbital, chloral hydrate, morphine, physostigmine, Δ9 THC, hemicholinium-3 and oxotremorine, drugs which decrease ACh turnover and release, caused a reduction in choline uptake. Conversely, administration of pentylenetetrazol, atropine, scopolamine, and haloperidol, drugs which increase ACh turnover and release, caused an increase in choline uptake . These findings support the proposal that sodium-dependent high affinity choline uptake can be used as a relative measure of the activity of cholinergic neurons . 相似文献
9.
A particulate factor of rat liver is described which interconverts three forms of rat liver cytosolic tyrosine aminotransferase with no alteration of enzyme activity. The factor appears to be a heat- and pH-sensitive lysosomal protein. The interconversion process is stimulated by 2.5 mM MgCl2 and 2.5 mM ATP. Asparate aminotransferase multiple forms are also susceptible to interconversion by the lysosomal factor. The properties of the factor explain several anomalous effects of manipulation on the tyrosine aminotransferase forms which have been reported in the literature and implicate the form interconversion in the degradation of tyrosine aminotransferase. 相似文献
10.
Incorporation of C14 Leucine was determined or in isolated mitochondria and microsomes of rat brain and liver after acute or chronic ethanol administration .The protein synthesis in mitochondrial and microsomal preparation was inhibited respectively by chloramphenicol and cycloeximide, specific inhibitors for the two systems tested. The experimental data demonstrate that the protein synthesis in both systems, mitochondrial and microsomal, is strongly affected only after chronic treatment which produces significant activation at the mitochondrial and microsomal level in the liver and an inhibition on the same systems of the brain.The data for protein synthesis instead show strong inhibition after acute administration, except for brain mitochondria, which are practically unaffected, while after chronic treatment no significant alterations are observed. 相似文献
11.
E Ambellan 《Biochemical and biophysical research communications》1974,57(2):520-525
Glucocorticoid binding to certain cell particles of rat liver and thymus following treatments and consists in part of a very “tight binding” that resists hot and cold perchloric acid extractions. This binding is found in thymus nuclei and in liver cytoplasmic particles, but not in liver nuclei nor in thymus mitochondria or microsomes. The existence of “tight binding” coincides with the ability of the same particles to bind free corticoid directly in incubations . The difference in the cellular location of this binding suggests that different methods of glucocorticoid activation exist in the anabolic target tissue, liver, and the catabolic target tissue, rat thymus. 相似文献
12.
The effect of beta-naphthoflavone on rabbit liver protein synthesis, and on the induction of cytochrome P-450 LM4 mRNA 总被引:1,自引:0,他引:1
K L Nash J Y Chiang A W Steggles 《Biochemical and biophysical research communications》1981,100(3):1111-1117
A single injection of β-naphthoflavone dispersed in corn oil causes significant changes in rabbit liver polysome and polysomal poly(A+)mRNA driven protein synthesis. The changes occur between 6–18 hr and 30–36 hr after the injection. Our data indicate that the first effect is due to the β-naphthoflavone and the second effect is due to the oil vehicle. translation of rabbit liver polysomes obtained from treated rabbits followed by specific immunoprecipition and gel electrophoresis, showed that maximal levels of translatable cytochrome P-450 LM4 occurred 18–24 hr after β-naphthoflavone treatment. 相似文献
13.
Pretreatment of rats with clorgyline, a selective inhibitor of MAO-A, significantly inhibited the deamination of intraventricularly administered serotonin (5-HT) and 5-methoxytryptamine (5-MT), but not phenylethylamine (PEA). Pretreatment with d, l-deprenyl, a selective inhibitor of MAO-B, significantly inhibited the deamination of all three substrates. Brain and liver homogenates from rats pretreated with clorgyline showed a decreased ability to deaminate () 5-MT and 5-HT, but not PEA. Homogenates from animals pretreated with d,l-deprenyl showed a decreased capacity to deaminate PEA, but not 5-MT or 5-HT. Clorgyline, when added to brain and liver homogenates, selectively blocked the deamination of 5-MT and 5-HT, but not PEA, whereas, d,l-deprenyl blocked the deamination of PEA without affecting that of 5-MT or 5-HT. In addition, 5-MT was found to be 100 X more potent than PEA at inhibiting the deamination of 5-HT. These findings suggest that 5-MT and 5-HT are favored substrates for MAO-A and . However, , significant amounts of 5-MT and 5-HT can also be deaminated by MAO-B. 相似文献
14.
The isolated myenteric plexus-longitudinal muscle of the guinea-pig ilem (GPI) was used as testsystem to study the influence of chronic morphine treatment on the levels of enkephalins, β-endorphin and some of its fragments. The peptides were assayed by means of a combination of high pressure liquid chromatography and radioimmunoassays. It was found that the levels of methionine- and leucine-enkephalin and β-endorphin were not altered by chronic morphine treatment of guinea-pigs nor in GPI exposed to morphine . However, the levels of some β-endorphin fragments i.c. γ-endorphin and des-tyrosine-γ-endorphin were elevated after morphine treatment and respectively. It is suggested that β-endorphin and its fragments are involved in homeostatic processes during development of opiate tolerance. 相似文献
15.
The effects of glucagon and insulin administration on hepatic mitochondrial Ca2+ uptake were compared with the effects of these hormones when they were added directly to the perfused liver. Glucagon administration increased mitochondrial calcium uptake both and in the perfused liver. In contrast, while injection of insulin into rats stimulated, addition of insulin to the perfusate, inhibited Ca2+ uptake. Cyclic AMP, when added to the perfusate, also increased the uptake of Ca2+ by mitochondria, subsequently isolated. The possible implications of the results are discussed. 相似文献
16.
The intracellular distribution of 115cadmium was determined following a pulsed exposure to the metal. The uptake and disappearance of label from rat liver nuclei was correlated with the appearance of a cytoplasmic Cd-binding protein. By coupling - experiments it was shown that unspecifically bound cadmium is free to enter the nucleus while specifically bound cadmium remains in the cytoplasm. 相似文献
17.
Marja Hollemans Ronald Oude Elferink De Groot Anneke Strijland Joseph M. Tager 《生物化学与生物物理学报:生物膜》1981,643(1):140-151
The volume of the lysosomal compartment in cultured human skin fibroblasts was estimated from the distribution between the cells and the medium of tracer amounts of labelled methylamine and chloroquine, which accumulate in the lysosomes, 2,2-dimethyloxazolidine-2,4-dione, which accumulates in the soluble cytoplasmic compartment relative to the lysosomes, and sucrose, which is excluded by the cells. In a foetal fibroblast line, the fractional volume of the lysosomal compartment was (). In fibroblasts from a patient with the I-cell disease, the fractional volume was 0.15.The fractional volume of the lysosomal compartment was used to calculate the intralysosomal pH from the accumulation of the weak bases in the cells. The mean value obtained was ().In fibroblasts incubated with various concentrations of chloroquine, the fractional volume of the lysosomal compartment and the accumulation of chloroquine in the cells were used to calculate the concentration of chloroquine in the lysosomes. The intralysosomal concentration increased from 3 to 114 mM as the extracellular concentration increased from 1 to 100 μM. Concomitantly, the intralysosomal pH increased from 5.3 in the absence of chloroquine to 5.9 in the presence of 100 μM chloroquine. A similar increase in intralysosomal pH could be calculated in fibroblasts incubated with different concentrations of ammonia. 相似文献
18.
Two populations of L5178Y murine leukemic cells, maintained by different methods, were studied for their implantation ability in BDF1 mice. Implantation ability was measured by number of tumor nodules formed, liver weight, and day of death of the animal. 1) Cells from a population grown for 10 years had no implantation ability; i.e., no tumor nodules were formed when injected into the tail vein. After 30 days of growth in the peritoneal cavity of BDF1 mice, these same cells were injected into the tail vein and 10 days later had produced over 200 liver tumor nodules. When cells taken from these tumors were recultured for 60 days , they lost the acquired implantation ability, but regained it after another single peritoneal passage. 2) L5178Y murine leukemic cells grown for six years in ascites tumor cells were extremely tumorigenic; over 200 tumor nodules appeared in the liver after tail vein injection. These cells were not rendered less tumorigenic and did not lose their implantation ability by culturing for 60 days. The results suggest that implantation ability is a property of the cell's growth environment; furthermore, they have strong implications for the and manipulation of this property. 相似文献
19.
Effect of aphidicolin on viral and human DNA polymerases. 总被引:9,自引:0,他引:9
DNA polymerases induced by Herpes simplex and Vaccinia viruses are inhibited by aphidicolin and this inhibition is probably the basis of its antiviral activity . Its possible clinical use is however hampered by the concomitant effect on human replicative DNA polymerase α. The inhibition of human α-polymerase is reversible both and and the changes in the rate of incorporation of thymidine into DNA, following treatment with aphidicolin for a generation time, indicate the likely synchronization of the cells due to this agent. DNA polymerase β, which has recently been shown to carry out repair synthesis of damaged nuclear DNA, is not inhibited by aphidicolin either on suggesting that the drug could allow a rapid and simple evaluation of DNA repair synthesis due to DNA polymerase β. 相似文献
20.
Some opiates with morphinan- and benzomorphan-structures possess affinities for neuroleptic receptors as revealed by their abilities to compete with 3H-spiroperidol for common binding sites in rat striatum (IC50 in the range between 10?6 and 10?5M). The binding of these opiates to neuroleptic receptors appears to be of pharmacological significance, since studies in mice revealed a small but significant displacement of spiroperidol by high doses of the opiate antagonist levallorphan from specific binding sites in the striatum. In addition, there exists some correlation between the ability of opiates to bind to neuroleptic receptor sites and their potency to evoke “bizarre behavior” in rats . In contrast, a wide variety of other opiates having morphine-, morphinone- or oripavine-structure showed no affinity for neuroleptic binding sites (IC50 greater than 10?4 M). Of the opioid peptides (methionine-enkephalin, leucine-enkephalin and β-endorphin) none has an affinity for neuroleptic binding sites. A variety of other peptides were also investigated but did not interfere with spiroperidol binding. Only ACTH showed a moderate affinity for neuroleptic binding sites. 相似文献