Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound   Target Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound

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Query: EC:3.1.3.5 (5'-nucleotidase)
3,167 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

The lumenal plasma membrane has been isolated from transitional epithelial cells (urothelium) lining the urinary bladder in sheep by a modified technique involving treatment with hypotonic thioglycolate. The isolated membranes, like those in situ, are distinguished morphologically by arrays of hexagonal particles (in plague regions) separated by smooth interplaque regions. These plaque regions, specifically, can be isolated from the lumenal plasma membrane. Of the proteins constituting the lumenal plasma membrane, five were found to characterize the plaque regions and, in particular, the 33,000-dalton species appears to be most heavily concentrated in the sodium dodecyl sulfate-polyacrylamide gel pattern of the isolated plaque regions. Lipid analyses showed that there are approximately 0.93 mg of phospholipid and 0.27 mg of cholesterol for each milligram of protein, giving a value of 55% lipids and 45% proteins for the composition of the lumenal plasma membrane. The total sialic acid content was measured to be approximately 0.038 micronmol/mg protein for the plasma membrane. Several plasma membrane marker enzymes were found to be associated with the lumenal plasma membrane fraction, but only the 5'-nucleotidase activity was found to be further enriched in the plaque region fraction. Amino acid analysis of the intrinsic proteins of the plaques indicated a polarity index of 45%.
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PMID:Isolation and characterization of the urothelial lumenal plasma membrane. 19 35

Stable cultures of mononuclear phagocytes from carrageenan-induced granulomas in mice have been established after enzymatic dispersion of these lesions. The cells can be maintained for up to 3 wk without division in serum-free media. The mononuclear phagocytes were identified by several criteria. The cells are adherent, phagocytic, contain lysosomal acid hydrolases at high specific activities, secrete lysozyme, and bind soluble aggregates of IgG. The activities of 5'-nucleotidase and leucine aminopeptidase in the cultured granuloma cells showed that they resembled macrophages from thioglycollate-stimulated mice but not unstimulated macrophages in these respects. Supernates from the cultured granuloma cells contain factor(s) which induce the proliferation of thymocytes; the release of such factors by the cells is stimulated by lipopolysaccharide.
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PMID:Mononuclear phagocytes from carrageenan-induce granulomas. Isolation, cultivation, and characterization. 67 Aug 87

Mouse resident peritoneal macrophages display sufficient 5'-nucleotidase activity to hydrolyze 58 nm AMP/min per cell protein. This activity increases approximately 163 nm AMP/min per mg after 72 h in culture. The enzyme is renewed in unstimulated cells with a half-time of 13.9 h. The activity is not reduced by treatment of intact cells with a variety of proteolytic enzymes, including trypsin, pronase, urokinase, and plasmin. Cells obtained from an inflammatory exudate have diminished or absent levels of enzyme activity. Endotoxin-elicited cells display enzyme activitiy of 20.9 nm AMP/min per mg, while thioglycollate-stimulated macrophages have no detectable activity. The reduced level of activity in endotoxin-stimulated cells is due to their elevated rate of enzyme degradation, with a half-time of 6.9 h. Their rate of enzyme synthesis is essentially normal. No evidence for latent enzyme activity could be obtained in thioglycollate-stimulated cells, nor do these cells produce any inhibition of normal cell enzyme activity. Serum deprivation reduces the enzyme activity of resident cells to about 45% of control activity. These conditions do not significantly affect the rate of enzyme synthesis, but again are explainable by an increase in the rate of enzyme degradation. Pinocytic rate is elevated in endotoxin-stimulated cells which show a more rapid rate of enzyme degradation than unstimulated cells do. However, in serum-free conditions, the rate of enzyme degradation is doubled with no change in the pinocytic rate of the cells.
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PMID:5'-Nucleotidase activity of mouse peritoneal macrophages. I. Synthesis and degradation in resident and inflammatory populations. 100 5

The binding of 125I-labeled thrombin to rat peritoneal macrophages isolated 20 h after the ip injection of thioglycollate broth or lipopolysaccharide decreased to 20% of the value found in resident macrophages due to a decrease in the number of receptors. The binding returned to normal values within a week after the injection. The decline parallelled more or less the Vmax for the 5'-nucleotidase activity. This decrease in the binding of thrombin could not be explained by an immigration of monocytes into the peritoneal cavity, since the binding of 125I-labeled alpha 2-macroglobulin-trypsin complex increased 4.5-fold in the same cell population due to an increase in the number of receptors, and blood monocytes do not bind alpha 2-macroglobulin-trypsin complex. The increase in the binding of alpha 2-macroglobulin-protease complex parallelled an increase in the incorporation of glucosamine, although the latter did not increase to the same extent. Engulfment of plasma membrane after phagocytosis did not result in a decreased binding of thrombin, but preincubation at 37 degrees C with concanavalin A caused a minor reduction in the binding. There was a positive correlation between the binding of alpha 2-macroglobulin-trypsin complex and the fraction of polymorphonuclear leukocytes in the peritoneal exudate and a negative correlation between the binding of thrombin and the fraction of polymorphonuclear leukocytes in the exudate, when the inflammation was induced by a milder stimulus, sterile NaCl, indicating a common signal for the polymorphonuclear leukocyte chemotaxis and the macrophage differentiation.
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PMID:In vivo inflammatory stimulation induces a transient change in the binding of thrombin to rat peritoneal macrophages. 131 45

Macrophages (m phi s), important cells in host resistance, undergo a series of biochemical changes during their progression from the resident to the fully activated stage. Both resident and inflammatory m phi s are characterized by some unique properties. In the present study, female BALB/c mice were prenatally treated with 8 mg/kg body weight of chlordane, a cyclodiene poly-chlorinated hydrocarbon that appears to reduce immunocompetence by selectively impairing m phi function. Therefore, we examined functions in m phi s from chlordane-treated mice that had been stimulated with thioglycollate. The 5'-nucleotidase activity, present in high levels in resident m phi s but low levels in inflammatory m phi s was elevated in resident m phi s from vehicle-exposed animals. Conversely, inflammatory m phi s from these animals showed significantly diminished levels of this function. Moreover, chlordane-exposed m phi s, regardless of whether they were resident or inflammatory, exhibited decreased 5'-nucleotidase responses. When a second function, transferrin receptor binding, was analyzed, vehicle-treated inflammatory m phi s displayed high levels of activity whereas the resident m phi s showed very little transferrin binding. However, both resident and inflammatory m phi s from the chlordane-exposed group demonstrated transferrin binding activity similar in magnitude to that of the vehicle-treated inflammatory m phi s. Finally, two-dimensional polyacrylamide gel electrophoresis analysis of m phi s from chlordane-exposed mice have characteristics of normal m phi s that have advanced to the inflammatory stage.
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PMID:In vivo prenatal chlordane exposure induces development of endogenous inflammatory macrophages. 156

The phenotype of three ectoenzymes was determined for murine resident peritoneal macrophages, macrophages elicited in vivo by treatment of mice with thioglycollate, Corynebacterium parvum or pyran, and for resident macrophages activated in vitro by treatment with lymphokine. The relationship of these biochemical markers to macrophage antiviral and anti-tumor activity was established. Thioglycollate-elicited macrophages showed a unique ectoenzyme phenotype, with increased leucine aminopeptidase and alkaline phosphodiesterase I activity and markedly reduced 5'-nucleotidase activity as compared with resident macrophages. Thioglycollate-elicited macrophages exhibited extrinsic antiviral activity against herpes simplex virus but did not show anti-tumor activity. Another ectoenzyme phenotype was shared by macrophages elicited in vivo by treatment of mice with the immunomodulators or in vitro by treatment with antigen-specific lymphokine. These macrophage populations showed increased levels of leucine aminopeptidase but reduced levels of both 5'-nucleotidase and alkaline phosphodiesterase. This ectoenzyme phenotype was associated with the acquisition by the macrophages of selective anti-tumor activity. There appear to be clear distinctions in biochemical markers and functional properties among macrophages activated by different mechanisms.
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PMID:Changes in macrophage ectoenzymes associated with anti-tumor activity. 625 Nov 33

Macrophages have been obtained from the peritoneal cavities of C57BL/6 mice following treatment with C. parvum, MVE-2, mineral oil, or thioglycollate. Cell populations were primarily composed of mononuclear phagocytes as determined by a latex bead uptake assay. Macrophages obtained from C. parvum or MVE-2 were activated as judged by enhanced cytostatic activity against two tumor cell target lines. Thioglycollate-elicited macrophages demonstrated much lower cytostatic ability. Rats were immunized with activated MVE-2 macrophages. Hybridomas were prepared by fusion with a non-secreting myeloma cell line followed by cloning. Cell supernates were selected on the basis of binding to activated but not elicited macrophages. The monoclonal antibody produced has been characterized by flow cytometry. The antibody does not react with syngeneic erythrocytes, thymocytes, or spleen cells. Reaction with thioglycollate macrophages is very low. Alternatively, intense binding is found on activated macrophages. This antigen which accompanies macrophage activation for tumor cell cytostasis is designated as macrophage activation antigen-1 (MAA-1). Several important physiological changes accompany the process of macrophage activation. For example, activated macrophages demonstrate enhanced microbicidal, phagocytic, secretory, and tumoricidal activity (for reviews see refs. 1,2). Concommitant alterations in cell surface properties have been observed. These include: (a) changes in surface morphology and spreading (3-5), (b) altered lipid and protein content (6,7), (c) decreases in 5'-nucleotidase activity and alkaline phosphodiesterase (8), increases in leucine aminopeptidase (8), decreases in mannose receptors (11,12), and antigen F4/80 (11), (d) increases in Ia antigens (11,12), and (e) increased tumor cell binding (13). These structural and functional modifications indicate that activated macrophages represent a unique class of functionally differentiated cells (9). Antigenic modifications accompanying macrophage differentiation are of special interest. Markers for specific macrophage classes might be useful in defining differentiation pathways, dissecting type-specific functional activities such as tumor cytotoxicity, and providing a means to identify macrophage subsets in heterogeneous cell populations. In the present work we have taken the first step in this direction by defining a cell surface macrophage activation antigen.
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PMID:Characterization of a monoclonal antibody defining a macrophage activation-specific cell surface antigen. 674 39

The administration of the bone-seeking isotope, 89Sr, to mice results in severe monocytopenia without any apparent effect on the numbers of resident peritoneal macrophages (Mphi). An explanation for this dichotomy was sought by determining whether the residual blood monocytes were still an effective source of Mphi after 89Sr treatment. Stem cell enumeration showed that a 90% fall in bone marrow macrophage colony-forming cells after 89Sr was accompanied by a 10-fold rise in splenic M-CFC. Splenectomy performed before 89Sr treatment, however, resulted in little additional monocytopenia and had no affect on the numbers of resident peritoneal Mphi even when sampling was extended to 31 days, an interval beyond the accepted half-time for peritoneal Mphi. Intraperitoneal injections of thioglycollate or Corynebacterium parvum elicited few or no monocyte-Mphi during respective intervals of 4 and 7 days. Elicitation with thioglycollate was attempted in tritiated thymidine-labeled mice 26 days after 89Sr. Four days later only a 2-fold increase in labeled peritoneal Mphi was found in the 89Sr-treated mice compared with a 150-fold increase in the controls. Studies of the ectoenzymes 5'-nucleotidase, alkaline phosphodiesterase I, and leucine aminopeptidase in such elicitation experiments suggested that the observed changes in activities reflected the direct stimulation of resident Mphi rather than monocyte immigration. Overall, the results indicate that treatment with 89Sr distinguishes two large populations of Mphi on the basis of their dependence on bone marrow. Mphi of inflammation reflect the monocytopenia and are severely and rapidly depleted by such treatment. The maintenance of resident type Mphi, on the other hand, appears to be independent of both the state of the bone marrow and the level of monocytes in the blood.
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PMID:Differential effects of chronic monocyte depletion on macrophage populations. 688 84

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