Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:3.1.3.1 (alkaline phosphatase)
 47,916 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

The effect of one-week exposure to sodium butyrate on HeLa S3 cell cultures was studied with special regard to influence on prekeratin synthesis, by comparison to cultures similarly treated with the known proliferation inhibitor hydroxyurea, and not treated. Like hydroxyurea, sodium butyrate inhibited cell proliferation to a considerable degree, but accounted additionally for an increase in membrane-bound alkaline phosphatase activity, cellular prekeratin synthesis, tonofilament number, and filament bundle formation. These phenomena unequivocally indicate that sodium butyrate acted as a specific stimulator of Hela (epithelial) cell differentiation. Similar differentiation phenomena can be observed during early spontaneous keratinization of the stratified horny epithelium.
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PMID:Influence of sodium butyrate on HeLa cell morphology and proliferation. 241 98

We have devised an enzyme-linked immunosorbent assay (ELISA) to quantitate fluid phase terminal complement pathway activation. Upon activation to form C5b-9, terminal complement components express neoantigens not present in the unassembled individual components. Expression of one of these neoantigens occurs at the step of C9 activation. C9 neoantigen is present in fluid phase SC5b-9 complexes, membrane-bound MC5b-9 complexes, and in in vitro polymerized C9. Under physiologic conditions, the presence of C9 neoantigen indicates that the terminal complement pathway is activated through the terminal component C9. In our assay for C9 neoantigen, we used rabbit antiserum to polymerized C9 rendered specific for C9 neoantigenic determinants by serial absorption with human serum, human C9, and other terminal complement components bound to Sepharose. Using the IgG from this antiserum, we devised a sandwich ELISA to bind SC5b-9 from solution onto polystyrene plates. The ELISA plates were developed with the use of goat antiserum to native C9 epitopes followed by a swine anti-goat IgG-alkaline phosphatase conjugate. Quantitation of SC5b-9 in solution was performed by comparing sample OD to a standard curve generated with human SC5b-9 that was purified from zymosan-activated serum. The assay was sensitive to as little as 100 ng of SC5b-9/ml and should be useful for screening plasma, serum, cerebrospinal fluid, or other biological fluids for the presence of terminal complement pathway activation.
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PMID:Quantitation of activation of the human terminal complement pathway by ELISA. 241 46

NKI/C-3 and NKI/black-13 are monoclonal antibodies recognizing different epitopes on a melanoma-associated antigen that is preserved after fixation in formalin and embedding in paraffin in virtually all melanoma tissues. The antigen, a predominantly cytoplasmic vesicle membrane-bound heterogeneous glycoprotein of 25-110 X 10(3) daltons, was shown to be a single 25 X 10(3) dalton polypeptide when incorporation of N-linked carbohydrates was inhibited by tunicamycin. The antigen was measured in a double determinant enzyme immunoassay (DDEIA) using NKI/C-3 as catcher antibody. Results from in vitro experiments indicated that the antigen is actively shed from living cells. In sera from melanoma patients with a small tumor burden, the antigen concentrations were in the range of those of controls (0-22 U/ml). Significantly increased values (33-600 U/ml) were found in sera from patients with a moderate or large tumor burden. The antigen concentrations in sera from patients with multiple metastases of other tumors were within the range of controls. Several sera from patients with multiple metastases of colon, pancreatic, and stomach carcinoma, however, contained increased antigen concentrations (45-80 U/ml). These results correspond with the reactions of NKI/C-3 in tissue sections of some malignancies other than melanoma. During the follow-up of melanoma patients the concentrations of circulating antigen correlated with tumor progression. The predictive value of the NKI/C-3 assay was no better than determination of serum lactate dehydrogenase, alkaline phosphatase or gamma glutamyl transferase activity.
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PMID:Circulating melanoma-associated antigen detected by monoclonal antibody NKI/C-3. 243 Jul 6

An improved aqueous two-phase polymer method has been developed for the isolation of sperm plasma membranes by manipulating various parameters that influence markedly the purity as well as yield of the membrane. The method consists of hypotonic shock of intact spermatozoa with 1.25 mM EDTA to dissociate the plasma membrane and dispersion of these cells to a two-phase polymer system consisting of 5.5% 252-Kd dextran and 4.2% 20-Kd polyethylene glycol prior to centrifugation at 9700 X g for 30 min when the two polymer phases are separated; the membrane fraction sediments at the interphase. The resulting membrane fraction was purified further by repeating the two-phase fractionation step. The yield of the membranes was approx. 35-40%, based on the recovery of the membrane-bound marker enzymes alkaline phosphatase and 5'-nucleotidase. The isolated membranes showed a high degree of purity as evidenced by phase contrast and electron microscopic studies and analyses of marker enzymes characteristic of cellular organelles. The yield and purity of the membranes have been found to be markedly dependent on the conditions of the hypotonic shock, obtained as a function of, EDTA concentration and on the molecular sizes of the dextran and polyethylene glycol that constitute the two-phase polymer system, as well as on the centrifugal force used for the sedimentation of the membrane.
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PMID:Factors influencing the yield and purity of goat sperm plasma membranes isolated by means of an aqueous two-phase polymer system. 244 37

A panel of 14 monoclonal antibodies (McAb) against hematopoietic cell surface antigens was applied on mononuclear blood or bone marrow cells from 40 cases of acute leukemia in order to compare immunoenzymatic staining (IE) (alkaline phosphatase) of fixed cells with immunofluorescence staining (IF) of unfixed suspended cells. According to the immunological results, 25 cases were phenotyped as ALL and 15 cases as AML. Cases with blast crisis secondary to chronic myelogenous leukemia (CML-BC) were not represented in this series. In all ALL cases the two methods gave an identical antigenic distribution. In 20 our of 21 cases of non-T-cell ALL, a B-cell progenitor origin was demonstrated by a positive staining reaction with the anti-CD19 McAb AB1 or HD37, and in 10 cases additionally with the anti-CD20 McAb B1 or 1F5. In contrast to the results obtained with IF, IE revealed a poor preservation of the AB1 epitope on CD19, whereas the HD37 epitope was equally well demonstrated by both methods. In 15 cases of AML the distribution of positive versus negative cells with IE or IF was identical for all McAb except J5 (anti-CALLA) (CD10) and B1 (CD20). Thus, 10/15 AML cases expressed CALLA with IE compared to 2/15 with IF. The corresponding figures for B1 were 5/15 and 0/15, respectively. Accordingly, normal myeloid precursor cells were CALLA-positive with IE but negative with IF. The discrepancy probably reflects the fact that, whereas both intracytoplasmatic and membrane-bound antigens are exposed in IE, only the latter are in IF. If the alteration of antigenic accessibility after fixation is considered, IE can safely be used for immunophenotyping of acute leukemia.
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PMID:Immunological typing of acute leukemias: immunoenzymatic staining of fixed cells compared with immunofluorescence staining of unfixed cells in suspension. 245 10

The clinical course and hematologic changes of three dogs with lymphocytosis of cells morphologically resembling large granular lymphocytes are presented. Hemograms from all dogs showed leukocytosis with marked lymphocytosis. Lymphocytes were characterized by abundant basophilic cytoplasm containing distinct granules which varied in size and number. Electron microscopically the granules were membrane-bound with an electron-dense core. Lymphocytes from one dog were positive for alkaline phosphatase activity, and lymphocytes from another dog were positive for alpha naphthyl butyrate esterase activity. Lymphocytes from one dog were positive for surface receptors for the crystalline fraction portion of gamma immunoglobulins.
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PMID:Lymphocytosis of large granular lymphocytes in three dogs. 246 45

Intramembranous localization of alkaline phosphatase (orthophosphoric monoester phosphohydrolase, alkaline optimum, EC 3, 1, 3, 1; AlPase) was observed biochemically in Bacillus megaterium KM grown in 1% polypeptone medium containing 0.5% NaCl at 37 degrees C under aerobic conditions and harvested at the latter logarithmic phase. AlPases from B. megaterium have been separated into soluble and membrane-bound forms by the centrifugation after cell disruption by sonication. The membrane-bound enzyme was further fractionated to two forms by phase separation using a non-ionic detergent, Triton X-114; one was successfully solubilized into the aqueous phase and the other remained in the Triton phase. Both AlPases of sonication- and Triton-solubilized forms were partially purified by gel filtration and anion-exchange column chromatographies. Their molecular weights were different (52,000 for soluble and 66,000 for Triton-solubilized forms) and the Vmax of the sonication-solubilized enzyme (227 nmol/min/mg protein) was 11-fold higher than that of the Triton-solubilized one although similar Km values (1.7 and 2.3 mM) were observed. Optimum pH of these enzymes tended to shift to a neutral range during the purification steps. These results suggest the multiplicity of AlPase anchoring to the membranes; 1) sonication-solubilized form which may be buried within the membrane lipids by its hydrophobic peptide and solubilized by the cell disruption, 2) detergent-solubilized form which may be bound loosely to the membrane by its hydrophobic domain and solubilized due to the amphiphilicity of enzyme protein, and 3) insolubilized form which may be bound fast to the membrane by its strong hydrophobicity and also have the function of enzymatic ability.
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PMID:[Biochemical studies on intramembranous localization of alkaline phosphatase in Bacillus megaterium KM]. 251 45

In order to investigate the membrane activities underlying development of neural cells, a histochemical localization of Ca2+-ATPase, Mg2+-ATPase and alkaline phosphatase (AlPase) activities in the rat cerebellar cortex during postnatal development was carried out. In the developing cerebellar cortex, ATPase activity was mainly associated with the plasma membranes of Purkinje and granular cells. This activity appeared in the immature Purkinje cells at birth and was proportionally increased throughout postnatal development. It was observed that the ATPase activity of migratory granular cells during a critical period from 3 and 15 postnatal days was increased in a funicular pattern in the developing cerebellar cortex. Conversely, peak AlPase activity in the developing cerebellar cortex was localized in the proliferative external granular cells until 7 postnatal days. Apparently, these phosphatase activities were not present in Bergmann glial fibers during the course of granular cell migration. The present findings were taken to indicate that neuronal cells in the cerebellar cortex have acquired a membrane-bound ATPase which can participate in Ca2+ transport or ATP metabolism during the course of early postnatal development.
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PMID:Histochemical localization of Ca2+, Mg2+-ATPase of the rat cerebellar cortex during postnatal development. 252 32

The SecY protein is a membrane-bound factor required for bacterial protein export and embedded in the cytoplasmic membrane by its 10 transmembrane segments. We previously proposed a topology model for this protein by adapting the Manoil-Beckwith TnphoA approach, a genetic method to assign local disposition of a membrane protein from the enzymatic activity of the alkaline phosphatase (PhoA) mature sequence attached to the various regions. SecY-PhoA hybrid proteins with the PhoA domain exported to the periplasmic side of the membrane have been obtained at the five putative periplasmic domains of the SecY sequence. We now extended this method to apply it to follow export of the newly synthesized PhoA domain. Trypsin treatment of detergent-solubilized cell extracts digested the internalized (unfolded) PhoA domain but not those exported and correctly folded. One of the hybrid proteins was cleaved in vivo after export to the periplasm, providing a convenient indication for the export. Results of these analyses indicate that export of the PhoA domain attached to different periplasmic regions of SecY occurs rapidly and requires the normal functioning of the secY gene supplied in trans. Thus, this membrane protein with multiple transmembrane segments contains multiple export signals which can promote rapid and secY-dependent export of the PhoA mature sequence attached to the carboxyl-terminal sides.
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PMID:Export of Escherichia coli alkaline phosphatase attached to an integral membrane protein, SecY. 253 43

The subcellular localization of the microbicidal nicotinamide adenine dinucleotide phosphate (NADPH) oxidase and associated b-cytochrome was investigated in human neutrophils. In unperturbed neutrophils 85% of b-cytochrome and the major part of membrane-bound components of the NADPH oxidase co-sedimented with markers for specific granules and gelatinase. Using cytochrome b559 as a marker for membrane-bound components of the NADPH oxidase in quantitative studies we observed that, of the remaining 15%, the vast majority co-sedimented with latent alkaline phosphatase, a marker for a newly identified mobilizable intracellular compartment. Only a small fraction co-localized with the plasma membranes. Azurophil granules contained a protease activity which rapidly inactivated the NADPH oxidase components present in other membranes. Stimulation of the neutrophils with formyl-methionyl-leucyl-phenyl-alanine and leukotriene B4 which caused minimal degranulation of specific granules, resulted in translocation of b-cytochrome to the plasma membrane, concomitant with incorporation of alkaline phosphatase into the plasma membrane.
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PMID:Dual granule localization of the dormant NADPH oxidase and cytochrome b559 in human neutrophils. 254 92


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