scholarly journals Subcellular localization of oestrogen-induced uterine peroxidase

1976 ◽  
Vol 160 (2) ◽  
pp. 237-241 ◽  
Author(s):  
C R Lyttle ◽  
P H Jellinck
Keyword(s):  
Acid Phosphatase ◽  
Subcellular Localization ◽  
Succinate Dehydrogenase ◽  
Density Gradient ◽  
Peroxidase Activity ◽  
Sucrose Density Gradient ◽  
Urate Oxidase ◽  
Mitochondrial Marker ◽  
Membrane Bound

The distribution of oestrogen-induced peroxidase in the resuspended 8000g pellet of rat uterine homogenates was examined by centrifugation in a sucrose density gradient. Within 10h of treatment with oestradiol, peroxidase activity was found in a region devoid of catalase or urate oxidase (peroxisomal markers) which did not overlap the fractions containing succinate dehydrogenase (mitochondrial marker) or acid phosphatase (lysosomal marker). The induced uterine enzyme was localized in reticular membrane-bound vesicles with isopycnic density of 1.28g/ml from which it could be released by treatment with detergent.

scholarly journals Subcellular localization of ferritin and iron taken up by rat hepatocytes

1989 ◽  
Vol 262 (2) ◽  
pp. 685-688 ◽  
Author(s):  
J C Sibille ◽  
M Ciriolo ◽  
H Kondo ◽  
R R Crichton ◽  
P Aisen
Keyword(s):  
Acid Phosphatase ◽  
Cytochrome C ◽  
Subcellular Localization ◽  
Cytochrome C Oxidase ◽  
Density Gradient ◽  
Rat Hepatocytes ◽  
Sucrose Density Gradient ◽  
Gel Chromatography ◽  
Density Gradient Ultracentrifugation

The subcellular localization of ferritin and its iron taken up by rat hepatocytes was investigated by sucrose-density-gradient ultracentrifugation of cell homogenates. After incubation of hepatocytes with 125I-labelled [59Fe]ferritin, cells incorporate most of the labels into structures equilibrating at densities where acid phosphatase and cytochrome c oxidase are found, suggesting association of ferritin and its iron with lysosomes or mitochondria. Specific solubilization of lysosomes by digitonin treatment indicates that, after 8 h incubation, most of the 125I is recovered in lysosomes, whereas 59Fe is found in mitochondria as well as in lysosomes. As evidenced by gel chromatography of supernatant fractions, 59Fe accumulates with time in cytosolic ferritin. To account for these results a model is proposed in which ferritin, after being endocytosed by hepatocytes, is degraded in lysosomes, and its iron is released and re-incorporated into cytosolic ferritin and, to a lesser extent, into mitochondria.

scholarly journals Localization of α-L-Arabinofuranosidase and Acid Phosphatase in Mycelium of Sclerotinia fructigena

Microbiology ◽  
2000 ◽  
Vol 81 (1) ◽  
pp. 79-99 ◽  
Author(s):  
E. C. Hislop ◽  
Vivien M. Barnaby ◽  
Claire Shellis ◽  
F. Laborda
Keyword(s):  
Acid Phosphatase ◽  
Density Gradient ◽  
Azo Dye ◽  
Acid Treatment ◽  
Sucrose Density Gradient ◽  
Subcellular Fractionation ◽  
Soluble Form ◽  
Ammonium Tartrate ◽  
Electron Microscopes ◽  
Hyphal Wall

α-L-Arabinofuranosidase (AF) and acid p-nitrophenyl phosphatase (AP) were secreted by Sclerotinia fructigena grown in a liquid pectin/ammonium tartrate medium. ‘Gentle’ mechanical manipulation of mycelium solubilized most of the AF and much of the AP, while brief acid treatment considerably inactivated both enzymes. Both enzymes were present predominantly in a soluble form in homo-genates prepared for subcellular fractionation, but some particulate activity of both was recovered from a sucrose density gradient in a fraction which also contained mitochondria. Azo-dye techniques with appropriate 1-naphthyl derivatives as substrates and p-(acetoxymercuric) aniline diazotate as capturing agent produced similar staining patterns for both enzymes in the light and electron microscopes, but the distribution of β-glycerophosphatase activity as visualized by the Gomori technique was more variable. A proportion of the activity of the enzymes remaining after fixation was located between the plasmalemma and the hyphal wall, in vacuoles in the cytoplasm, and in spherosome-like bodies. Some evidence was obtained for structure-linked latency of both enzymes and for their secretion by a process of reverse pinocytosis.

scholarly journals Subcellular localization of NAD(P)H oxidase(s) in human neutrophilic polymorphonuclear leucocytes

1978 ◽  
Vol 176 (1) ◽  
pp. 175-178 ◽  
Author(s):  
D B Iverson ◽  
P Wang-Iverson ◽  
J K Spitznagel ◽  
L R DeChatelet
Keyword(s):  
Cell Membrane ◽  
Nadph Oxidase ◽  
Subcellular Localization ◽  
Density Gradient ◽  
Membrane Fraction ◽  
Sucrose Density Gradient ◽  
Human Neutrophils ◽  
Polymorphonuclear Leucocytes

NADH and NADPH oxidase activities in a homogenate of human neutrophils co-sediment in a linear sucrose density gradient under either velocity or isopycnic conditions of centrifugation. The position of these activities in the gradient does not correspond to any known subcellular granule or to the cell-membrane fraction. These data suggest that the oxidase activities may reside in a unique granule that has previously not been recognized.

scholarly journals Free and membrane-bound polyribosomes in normal and Rauscher-virus-infected mouse spleen cells

1970 ◽  
Vol 119 (4) ◽  
pp. 749-756 ◽  
Author(s):  
J. Th. M. Burghouts ◽  
A. L. H. Stols ◽  
H. Bloemendal
Keyword(s):  
Density Gradient ◽  
Infected Mouse ◽  
Rat Liver ◽  
Sodium Deoxycholate ◽  
Sucrose Density Gradient ◽  
Spleen Cells ◽  
Ribonuclease Inhibitor ◽  
Rauscher Virus ◽  
Centrifugation Step ◽  
Membrane Bound

1. Free and membrane-bound polyribosomes and ribosomal monomers were isolated from normal and Rauscher-virus-infected mouse spleens by means of discontinuous sucrose density gradients. 2. The addition of ribonuclease inhibitor from rat liver was essential to protect these polyribosomes from degradation. To separate the smooth and rough membranes from ribosomal monomers an additional centrifugation step through a continuous sucrose density gradient was necessary. 3. After infection a marked increase in rRNA from both membrane-bound and free polyribosomes was observed. Treatment of the membrane-bound polyribosomes with sodium deoxycholate yielded only 80S particles even when ribonuclease inhibitor was added. 4. A striking feature of the infected spleen was the occurrence of large polyribosomes. Up to 40 monomers per polyribosome could be counted on electron micrographs.

scholarly journals Phototoxicity of protoporphyrin as related to its subcellular localization in mice livers after short-term feeding with griseofulvin

1981 ◽  
Vol 198 (1) ◽  
pp. 67-74 ◽  
Author(s):  
S Sandberg ◽  
I Romslo
Keyword(s):  
Acid Phosphatase ◽  
Oxidative Phosphorylation ◽  
Subcellular Localization ◽  
Succinate Dehydrogenase ◽  
Glutamate Dehydrogenase ◽  
Lysosomal Enzymes ◽  
Liver Mitochondria ◽  
Short Term ◽  
Rapid Accumulation ◽  
Uncoupling Of Oxidative Phosphorylation

In mice, feeding with griseofulvin leads to the rapid accumulation of protoporphyrin in liver mitochondria. When liver mitochondria from mice fed with griseofulvin for 2 days are exposed to irradiation (320-400 nm), uncoupling of oxidative phosphorylation followed by inhibition of respiration occurs at light doses above 3-5 kJ/m2. When combined preparations of mitochondria and lysosomes are irradiated, inactivation of enzymes occurs in the following order: succinate dehydrogenase greater than glutamate dehydrogenase greater than acid phosphatase greater than beta-glucuronidase. Qualitatively, the photodamaging effect of endogenously produced protoporphyrin is indistinguishable from that of externally added protoporphyrin. Quantitatively, however, when protoporphyrin is added externally, more protoporphyrin is taken up by lysosomes, and photoinactivation of the lysosomal enzymes is correspondingly more severe. The results are further evidence that porphyrin-induced photodamage is largely determined by the solubility properties of the porphyrins and the target structures [Sandberg & Romslo (1980) Biochim. Biophys. Acta 593, 187-195], and also that protoporphyrin-induced photodamage is essentially similar whether protoporphyrin is generated endogenously or added exogenously.

The Combining Ability of Glycoproteins IIb, IIIa and Ca2+ in EDTA-Treated Nonaggregable Platelets

1983 ◽  
Vol 50 (04) ◽  
pp. 848-851 ◽  
Author(s):  
Marjorie B Zucker ◽  
David Varon ◽  
Nicholas C Masiello ◽  
Simon Karpatkin
Keyword(s):  
Density Gradient ◽  
Combining Ability ◽  
Density Gradient Centrifugation ◽  
Gradient Centrifugation ◽  
Electrophoretic Pattern ◽  
Sucrose Density Gradient ◽  
Irreversible Loss ◽  
Sucrose Density Gradient Centrifugation ◽  
Crossed Immunoelectrophoresis ◽  
Triton X

SummaryPlatelets deprived of calcium and incubated at 37° C for 10 min lose their ability to bind fibrinogen or aggregate with ADP when adequate concentrations of calcium are restored. Since the calcium complex of glycoproteins (GP) IIb and IIIa is the presumed receptor for fibrinogen, it seemed appropriate to examine the behavior of these glycoproteins in incubated non-aggregable platelets. No differences were noted in the electrophoretic pattern of nonaggregable EDTA-treated and aggregable control CaEDTA-treated platelets when SDS gels of Triton X- 114 fractions were stained with silver. GP IIb and IIIa were extracted from either nonaggregable EDTA-treated platelets or aggregable control platelets with calcium-Tris-Triton buffer and subjected to sucrose density gradient centrifugation or crossed immunoelectrophoresis. With both types of platelets, these glycoproteins formed a complex in the presence of calcium. If the glycoproteins were extracted with EDTA-Tris-Triton buffer, or if Triton-solubilized platelet membranes were incubated with EGTA at 37° C for 30 min, GP IIb and IIIa were unable to form a complex in the presence of calcium. We conclude that inability of extracted GP IIb and IIIa to combine in the presence of calcium is not responsible for the irreversible loss of aggregability that occurs when whole platelets are incubated with EDTA at 37° C.

Characterization and Partial Purification of Heterodisperse Polysomal RNAs in Normal and Neoplastic Cells

1972 ◽  
Vol 58 (2) ◽  
pp. 71-94
Author(s):  
Ada Sacchi ◽  
Gianni Chinali ◽  
Susetta Pons ◽  
Michela Galdieri ◽  
Piero Cammarano
Keyword(s):  
Size Distribution ◽  
Density Gradient ◽  
Rat Liver ◽  
Sucrose Density Gradient ◽  
Partial Purification ◽  
Messenger Rnas ◽  
Alkaline Ph ◽  
Sedimentation Profile ◽  
Molecular Weights ◽  
Phenol Extraction

The size distribution of cytoplasmic messenger RNAs (m-RNA) has been studied in rat liver and in monodifferentiated cells (mouse reticulocytes and myelomas). It has been found that the RNA which exhibits a « rapid turnover » and a polydisperse profile of radioactivity is refractory to phenol extraction. This property has been exploited to selectively isolate m–RNA from the phenol residue by means of an extraction at an alkaline pH. The sucrose density gradient profiles of m–RNA isolated from monodifferentiated cells show monodisperse peaks having the sedimentation coefficients expected on the basis of the molecular weights of monocistronic messages for α and β chains of hemoglobin (reticulocytes) and L and H chains of immunoglobulin (myelomas). The sedimentation profile of cytoplasmic m–RNA associated with rat liver polysomes shows a much broader distribution, with sedimentation coefficients ranging from 8 S to 28 S.

scholarly journals Ribonucleic Acid Biosynthesis in Human Leukocytes

Blood ◽  
1966 ◽  
Vol 28 (2) ◽  
pp. 188-200 ◽  
Author(s):  
MARTIN J. CLINE
Keyword(s):  
Density Gradient ◽  
Ribonucleic Acid ◽  
Turnover Rate ◽  
Rna Synthesis ◽  
Gradient Analysis ◽  
Sucrose Density Gradient ◽  
Acid Biosynthesis ◽  
Total Load ◽  
Human Leukocytes ◽  
Particle Ingestion

Abstract Phagocytosis has profound effects on several aspects of the RNA metabolism of human leukocytes. The major changes induced by particle ingestion appear to be (1) an increased uptake of pyrimidine precursors from the suspending medium, (2) a contraction in the size of the nucleotide pool, (3) an accelerated rate of destruction of preexisting RNA, and (4) an increased rate of RNA synthesis. Sucrose density gradient analysis of the newly synthesized RNA suggests that several classes of RNA are involved in this process. The increased turnover rate of the nucleotide pool and of the cellular RNA of the leukocyte is proportional, within limits, to the total load of ingested particles.

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