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 predicted amino acid sequence of the alpha subunit of the rat liver mitochondrial ATP synthase has been obtained by sequencing a cDNA for the alpha subunit. Analysis of the sequence shows that it contains the A and B consensus sequences found in many nucleotide-binding proteins. Twelve amino acids of the rat liver alpha subunit differ from the sequence of the bovine heart alpha subunit; four of these involve differences in charge. The rat liver alpha subunit, from arginine 15 to the C-terminal proline 510, has been overexpressed in Escherichia coli using the alkaline phosphatase promoter (phoA) and leader peptide to direct the export of the expressed protein to the bacterial periplasm. By treating the cells with lysozyme, osmotic shock, and alkaline pH washes, the alpha subunit can be extracted in high yield (greater than 25 mg/liter) and in a high state of purity. The expressed alpha subunit remains soluble at pH 9.5 or greater and precipitates when treated with Mg2+ ions at low millimolar concentration. The bacterially expressed alpha subunit interacts with 2'(3')-O-(2,4,6-trinitrophenyl)adenosine 5'-triphosphate (TNP-ATP), resulting in a marked fluorescence enhancement upon binding. An enhancement of fluorescence is also observed upon the interaction of the alpha subunit with TNP-ADP. Preincubating the alpha subunit with 1.5 mM ATP significantly reduces the fluorescence enhancement seen with TNP-ATP. The alpha subunit binds TNP-ATP with an apparent Kd in the low micromolar range (1-5 microM) and binds TNP-ADP with an affinity at least 10-fold lower. This work shows that the rat liver alpha subunit can be overexpressed in E. coli to yield a large amount of functional protein. With the acquisition of the overexpressed alpha subunit, it is now possible to test the reconstitution of ATPase activity from a mixture of recombinant and rat liver-derived subunits and to test the formation of complexes by the overexpressed alpha and beta subunits of the rat liver F1-ATPase.
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PMID:Mitochondrial ATP synthase. cDNA cloning, amino acid sequence, overexpression, and properties of the rat liver alpha subunit. 213 25

Several immunohistochemical methods are now available for the staining of neoplastic cells in tissue sections. The authors have found that the alkaline phosphatase-anti-alkaline phosphatase (APAAP) method is sensitive and reliable. Murine monoclonal or nonmurine polyclonal antibodies can be used to label a variety of membranous and/or cellular constituents in tissues that have been routinely processed in a histopathology laboratory. The monoclonal antibody against leukocyte common antigen (CD45) can be used to differentiate hematologic from nonhematologic tumors. Monoclonal antibodies (L26, LN1, LN2, LN3, MB1, MB2) label B-cell lymphomas, whereas other monoclonal antibodies (UCHL1, MT1) more characteristically stain T-cell lymphomas. Polyclonal antibodies against CD3 specifically mark neoplastic cells from T-cell lymphomas and leukemias but as yet are not commercially available. Monoclonal antibodies Leu-M1 (CD15), Ber H2 (Ki-1; CD30), and LN2 label Reed-Sternberg cells from most cases of nodular sclerosis, mixed cellularity, and lymphocyte-depleted Hodgkin's disease. Monoclonal antibodies Mac 387, KP1 (CD68), and NP57 (antielastase), as well as polyclonal antibodies against lysozyme, help identify subtypes of acute myeloid leukemia and extramedullary myeloid cell tumors. Although there are now excellent reagents ready for use, there is still a significant need for more lineage-specific (particularly against CD epitopes) monoclonal antibodies capable of labeling neoplastic cells in paraffin-embedded tissue sections from patients with hematologic malignancies.
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PMID:Immunophenotyping of hematologic neoplasms in paraffin-embedded tissue sections. 218 Feb 77

We have investigated the relationship between monocytes/macrophages and bone metabolism with the in vitro study. In the present study monocytes/macrophages derived from rat bone marrow were harvested and their phenotype was examined. Collected cells exhibited many characteristics of monocytes/macrophages at various points. In addition, 1,25-dihydroxyvitamin D3(D3), one of the osteotropic hormones was found to alter the activities of acid phosphatase (AcPase) and lysozyme in the media and also stimulate alkaline phosphatase (AlPase) positive cells to increase in number.
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PMID:A harvesting method of monocytes/macrophages derived from bone marrow and their characteristics--special reference to the effect of 1,25-dihydroxyvitamin D3 on monocytes/macrophages. 231 98

Proximal tubular (PT) epithelial cells express MHC class II (Ia) antigens in immunologically-mediated renal injury. To study the role of PT as accessory cells, we generated several murine PT-like epithelial cell lines by transformation with origin-defective SV40 DNA. These transformed cell lines display typical alkaline phosphatase and gamma-glutamyl-transpeptidase enzyme activity, proliferation to epidermal growth factor (EGF) and sodium-dependent glucose uptake. Clonal lines of transformed tubular cells from both normal C3H/FeJ and autoimmune MRL-lpr mice do not constitutively express Ia antigens or mRNA for class II. However, stimulation with recombinant interferon-gamma(rIFN-gamma) induces Ia mRNA and surface product in the cell lines. These Ia-positive cells can process and present hen egg-white lysozyme (HEL) to antigen-specific Iak-restricted T cell hybrids. Unstimulated tubular cells do not express detectable IL-1 alpha, IL-1 beta, TNA-alpha, or IL-6 mRNA. However, stimulation with IL-1 alpha or LPS induces TNF-alpha transcripts. We conclude that these cell lines have characteristics most consistent with a proximal tubular origin. They also bear characteristics of accessory cells such as processing and presentation of antigen and TNF-alpha gene expression. We speculate that PT have the capacity to participate in the pathogenesis of immune renal injury.
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PMID:MHC class II, antigen presentation and tumor necrosis factor in renal tubular epithelial cells. 240 90

Exposure for 20 min of stationary phase cells of Salmonella typhimurium to a combined triple stress system (TSS) treatment comprising hypochlorite derived 5 ppm free available chlorine in solution acidified with 1% succinate (pH 2.5) and at a chill shock temperature of 5 degrees C resulted in symptoms of injury. Cells became sensitive to 40 micrograms/ml lysozyme, 50 micrograms/ml actinomycin D and 100 micrograms/ml ribonuclease B, to which control cells were resistant. Metabolic injury was indicated by reduction in colony forming ability of stressed cells on minimal salts glucose agar M9 medium. There was no detectable leakage loss of 260-280 nm-absorbing materials. This was also confirmed by assay of the cellular RNA material components. Loss of alkaline phosphatase activity was observed in the stressed cells. The intensity of induced cellular damage as measured by lysozyme sensitivity was greatest in the cells exposed to the complete TSS, followed by those stressed in 1% succinate at 5 degrees C, then 5 ppm chlorine at 5 degrees C and the singular chill shock stress at 5 degrees C, respectively. The magnitudes of cellular damage, however, were suggestive of synergistic interactions among the component stress factors of the TSS. The findings obtained indicated impairment of the structural integrity and functional capabilities of the permeability barriers and the inactivation of certain periplasmic enzymes. The resultant cumulative cellular damage from the TSS exposure may therefore enhance greater sensitivity of treated cells to subsequent stress factors.
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PMID:Mechanisms of triple stress-mediated damage in stationary phase cells of Salmonella typhimurium exposed to succinate-acidified hypochlorite system at 5 degrees C. 242

Human oesophageal submucosal glands may be regularly demonstrated by first exposing the oesophageal lumen to toluidine blue which reveals the duct ostia. Four types of cell were identified in the glands - mucous, subsidiary or serous, myoepithelial and oncocytes. The mucous cell contained neutral, sialated and sulphated mucins. The subsidiary cells held smaller amounts of neutral and sialated mucin, plus fucosyl residues. No lipids were detectable histochemically. ATP-ase and alkaline phosphatase were shown in the capillary endothelium. The duct epithelium showed some nonspecific esterase activity not sensitive to E 600. By immunoperoxidase techniques, the duct epithelium was shown to be rich in cytokeratin. The subsidiary cells contained lysozyme, CEA and pepsinogen. B lymphocytes composed most of the periductular lymphoid aggregates, although some T cells were found there and also intraepithelial and subepithelial in relation to the stratified squamous epithelium lining the oesophagus. Langerhans' cells were also demonstrated as intraepithelial by several techniques.
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PMID:Human oesophageal submucosal glands. Their detection mucin, enzyme and secretory protein content. 243 35

A sandwich ELISA for testing whether pairs of monoclonal antibodies recognize the same native antigenic site was developed. The assay was performed with murine anti-lysozyme monoclonal antibodies. A monoclonal antibody, adsorbed onto a microtiter plate, was used as a capture antibody for native lysozyme. After the reaction with the antigen a second monoclonal antibody, the test antibody, was added. The amount of bound antibody was quantitatively measured using rabbit anti-mouse immunoglobulin serum conjugated to alkaline phosphatase. The simultaneous binding of pairs of monoclonal antibodies to lysozyme was further substantiated by structural considerations.
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PMID:A simple ELISA for the classification of monoclonal antibodies according to their recognition of native epitopes. 244 86

A 62-year-old woman with chronic neutrophilic leukemia (CNL) is described. She presented in February 1988 for evaluation of leukocytosis of 3 years' duration with no complaint. Physical examination was normal. The leukocyte count was 20,100/microliters with 70% segmented neutrophils and 12% band forms. A myelogram showed marked myeloid hyperplasia and plasmacytosis (5.9%). Neutrophil alkaline phosphatase score, serum lysozyme and vitamin B12 levels were elevated. Cytogenetic analysis of the marrow aspirate showed normal karyotype, with no Philadelphia chromosome. Total serum protein (TP) was 7.5 g/dl with increased beta-globulin (23.5%), identified as monoclonal IgA kappa (3.3 g/dl) on immunoelectrophoresis. No activity of G-CSF was detected in the serum. A retrospective study revealed that the beta-globulin level was normal (6.3%, TP 6.9 g/dl) in 1980 and that it was slightly increased (11.6%, TP 7.0 g/dl) without leukocytosis (5,900/microliter) in 1981. In 1985, when leukocytosis obviously existed (9,900/microliter), the percentage of beta-globulin was increased to 17.5% (TP 7.2 g/dl). The possibility that monoclonal gammopathy preceded the leukocytosis must be admitted. On the basis of our observation, it is assumed that CNL and monoclonal gammopathy may be blood dyscrasias derived from a common precursor cell or that the immunological abnormality associated with monoclonal gammopathy may be implicated in the development of CNL.
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PMID:[Chronic neutrophilic leukemia associated with monoclonal gammopathy (IgA, kappa type)]. 250 2

Malignant fibrous histiocytoma (MFH) developed spontaneously in subcutaneous tissue of the head of a 15-month-old male Fischer 344 rat. The tumor was serially transplanted into syngeneic rats up to the 45th generation and was designated MFH-MT. Light and electron microscopic examinations revealed that the original and serially transplanted tumors were composed of an admixture of fibroblast-like and histiocyte-like cells arranged in a storiform pattern. Neoplastic cells gave positive reactions for acid phosphatase, alkaline phosphatase, nonspecific esterase, alpha-1 antitrypsin and lysozyme. The tumors transplanted into the lungs and cutaneous tissue of the tail had a mixed histologic appearance of storiform, pleomorphic, myxoid and giant cell types. Moreover sclerosing hemangioma-like and osteosarcoma-like structures were also found. MFH-MT grew well in athymic nude mice showing neoplastic proliferation of pleomorphic cells strongly positive for alpha-1 antitrypsin. Development of MFH-MT was significantly retarded by the two antitumor drugs tested. The retarded tumors consisted predominantly of fibroblast-like cells and abundant collagenic fibers, whereas histiocytic cells decreased in number.
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PMID:Morphologic characteristics of a transplantable tumor derived from a spontaneous malignant fibrous histiocytoma in the rat. 254 24

The following 10 enzymes were assayed in 187 amniotic fluid and maternal serum samples at 15-42 weeks of gestation: alkaline phosphatase, heat-stable alkaline phosphatase (only in amniotic fluid), acid phosphatase, alanine aminotransferase, aspartate aminotransferase, alpha-amylase, gamma-glutamyltransferase, creatine kinase, lactate dehydrogenase, and lysozyme. The normal reference ranges are reported for amniotic fluid and maternal serum enzymes, together with the abnormal values accompanying neural tube defects and EPH-gestosis. The determination of gamma-glutamyltransferase, heat-stable alkaline phosphatase and creatine kinase was found to be of appreciable diagnostic significance in clinical practice.
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PMID:Variation in some enzymes in amniotic fluid and maternal serum during pregnancy. 256 24


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