Showing papers on "Alkaline phosphatase published in 1972"
TL;DR: Gutman et al. as mentioned in this paper showed that the acid phosphatase of serum is reduced in metastatic carcinoma of the prostate by decreasing the activity of androgens through castration or estrogenic injections and that this enzyme is increased by injecting androgens.
Abstract: Carcinoma of the prostate gland is peculiarly favorable for endocrine investigation since frequent serial observations of the activity of phosphatases in serum were found to provide objective indices of activity of the neo/~i~m when the enzymes were increased in amount above normal. In the present paper data are given for the values of serum phosphatases in carcinoma of the prostate and in normal men. We shall demonstrate that the acid phosphatase of serum is reduced in metastatic carcinoma of the prostate by decreasing the activity of androgens through castration or estrogenic injections and that this enzyme is increased by injecting androgens. We have been unable to find previous observations indicating any relationship of hormones to carcinoma of the prostate gland. An enzyme capable of hydrolyzing phosphoric esters was discovered by Grosser and Husler (4) in intestinal mucosa and kidney. Robison (16) found that this enzyme was particularly high in activity in growing bone and cartilage and that its activity was greatest at pH 9 to 9.5. This ~alkaline phosphatase," was found by Kay (9) to be increased in the serum in certain bone diseases including metastasis of neoplasms to bone and later work has shown that among these conditions is carcinoma of the prostate. Davies (3) and Bamann and Riedel (1) discovered that there occurs in the spleen and kidney of swine and cattle, in addition to the alkaline phosphatase, a phosphatase with an activity maximum at pH 4.8. An enzyme believed to be identical with this "acid phosphatase" was found by Kutscher and Wolbergs (11) to be present in very large amount in the human prostate gland. This finding of great activity of acid phosphatase in the prostate gland was confirmed and extended to include prostatic cancer by Gutman, Sproul, and Gutman (7). The serum of certain patients with disseminated prostatic carcinoma was found by Gutman and Gutman (6) and Barringer and Woodard (2) to exhibit increased acid phosphatase activity. Robinson, Gutman, and Gutman'~I5) summarized the acid phosphatase activity levels of 44 patients with carcinoma of the prostate. They concluded that a marked rise in acid phosphatase in serum is associated with the appearance or spread of roentgenologically demonstrable skeletal metastases and implies dissemination of the primary tumor and thus is of unfavorable prognostic significance. METttODS AND MATERIALS
3,534 citations
TL;DR: Coarse powders of acid-insoluble matrix of diaphysis and calvarial parietal bone rapidly and consistently transformed fibroblasts into masses of cartilage and bone containing hemopoietic marrow.
Abstract: Coarse powders of acid-insoluble matrix of diaphysis and calvarial parietal bone rapidly and consistently transformed fibroblasts into masses of cartilage and bone containing hemopoietic marrow. The transformant was encapsulated by fibroblasts within 24 hr to form a plaque. Transformation was restricted to the central thicknesses of the plaque. Under the stated conditions the alteration of the phenotype, fibroblast to chondroblast, was an unstable transformation, whereas the phenotype change, fibroblast to osteoblast, was stable. The transformation occurred on a rigid timetable of sequences. Measurements of alkaline phosphatase activity and incorporation of radioactive sulfate, phosphate, and calcium were sensitive and quantitative assays for the appearance of the transformed products, cartilage and bone.
816 citations
TL;DR: A fluorometric method for assaying low levels of the enzyme alkaline phosphatase in seawater based on the hydrolysis of the monophosphate ester bond of 3-0-methylfluorescein phosphate is described.
Abstract: This paper describes a fluorometric method for assaying low levels of the enzyme alkaline phosphatase in seawater. The assay is based on the hydrolysis of the monophosphate ester bond of 3-0-methylfluorescein phosphate. This enzyme is synthesized by many microorganisms when phosphate becomes limiting. Alkaline phosphatase activity was detected in phytoplankton from the nutrient-impoverished surface waters of the subtropical Central North Pacific Ocean. The presence of naturally occurring phosphatase suggests that phosphorus may be limiting to phytoplankton growth in these waters. The phytoplankton in water samples lacking enzyme activity at the time of collection produced phosphatase within 1 to 2 days of incubation at in situ temperatures.
279 citations
TL;DR: The anthelmintic levamisole and one of its analogues, R 8231, were found to be very potent inhibitors of alkaline phosphatase from all tissues except intestine, and proved to be the most appropriate buffer.
240 citations
TL;DR: 2-Ethylaminoethanol supports the highest enzyme activity of any of the compounds tested, and Diethanolamine was shown to have many of the favorable characteristics required for a reference enzyme procedure.
Abstract: We have compared 23 compounds, with and without transphosphorylating properties, as buffer systems for human serum alkaline phosphatase activity, with p -nitrophenylphosphate as substrate. Relative enzyme activity in four representative buffers at near-optimal conditions was ethylaminoethanol > diethanolamine > 2-amino-2-methyl-1-propanol > carbonate. Transphosphorylation was demonstrated in the two buffers in which the enzyme was most active, ethylaminoethanol and diethanolamine. The optima for pH, buffer concentration, and substrate for these four systems were studied in detail. 2-Ethylaminoethanol supports the highest enzyme activity of any of the compounds tested. Diethanolamine was shown to have many of the favorable characteristics required for a reference enzyme procedure.
238 citations
Journal Article•
235 citations
TL;DR: The results suggest that increased collagen synthesis is a prerequisite for bone formation, but the significance of the developmental pattern of alkaline phosphatase in this process is unclear.
213 citations
TL;DR: Kinetic measurement of GGT activity offers a simple, sensitive, and direct means for distinguishing whether bone or liver is the source of increased serum alkaline phosphatase activity.
Abstract: Serum γ-glutamyl transpeptidase (GGT), leucine aminopeptidase, alkaline phosphatase, alanine aminotransferase, and aspartate aminotransferase activities were assayed in controls and in patients with liver, pancreatic, or bone disease. GGT activity was above normal in all forms of liver disease studied (viral hepatitis, cirrhosis, cholecystitis, metastatic carcinoma to liver, pancreatic carcinoma, liver granuloma, and acute pancreatitis). GGT more sensitively indicated hepatic disease than did alkaline phosphatase, much more so than did leucine aminopeptidase. GGT was disproportionately more active in relation to the transaminases in cases of intraor extrahepatic biliary obstruction; the reverse was true in cases of viral hepatitis. GGT activity was normal in children, adolescents, and pregnant women, and in cases of bone disease and renal failure. Kinetic measurement of GGT activity offers a simple, sensitive, and direct means for distinguishing whether bone or liver is the source of increased serum alkaline phosphatase activity. Activity was highest in obstructive liver disease.
197 citations
TL;DR: Since the product formed with poly(A) lacks 3'-hydroxyl ends, as measured with these exonucleases, the enzyme appears to convert linear molecules of polyriboadenylate to a circular form by the intramolecular covalent linkage of the 5'-phosphate end to the 3'-Hydroxyl terminus.
Abstract: An enzyme, purified 300-fold from Escherichia coli infected with bacteriophage T4, catalyzes the conversion of 5′-termini of polyribonucleotides to internal phosphodiester bonds. The reaction requires ATP and Mg++. For every 5′-32P terminus rendered resistant to alkaline phosphatase, an equal amount of AMP and PPi are formed. Various polyribonucleotides are substrates in the reaction; to date, the best substrate is [5′-32P]polyriboadenylate. With the latter substrate, no evidence of intermolecular reaction was obtained. However, the 5′-32P termini of poly(A) rendered resistant to alkaline phosphatase are also resistant to attack by RNase II, polynucleotide phosphorylase, and low concentrations of venom phosphodiesterase. Since the product formed with poly(A) lacks 3′-hydroxyl ends, as measured with these exonucleases, the enzyme appears to convert linear molecules of polyriboadenylate to a circular form by the intramolecular covalent linkage of the 5′-phosphate end to the 3′-hydroxyl terminus.
197 citations
TL;DR: Differences in substrate-activity relationships, and inhibition by l -phenylalanine and NaF provided additional support for the hydrolysis of phytate as an activity distinct from that promoting hydroleysis of p -nitrophenyl or 3-glycerophosphates.
183 citations
TL;DR: The apparent effect of certain drugs on serum GGT activity indicates the need for caution in interpreting the results of this test, and patients who had been treated with phenytoin and barbiturates were found to have elevated serum G GT activities without any other evidence of liver disease.
Abstract: Serum γ-glutamyl transpeptidase (GGT) activity correlates closely with the activities of alkaline phosphatase (ALP) and 5′-nucleotidase (5NT) in various forms of liver disease. Maximum elevations of all three enzyme activities are observed in diseases which particularly affect the biliary tract. Compared with the other two enzymes GGT is generally increased to a greater extent and is thus the most sensitive indicator of biliary-tract disease, while estimations of serum GGT are more reproducible than those of 5NT. However, a group of patients who had been treated with phenytoin and barbiturates were found to have elevated serum GGT activities without any other evidence of liver disease. The apparent effect of certain drugs on serum GGT activity indicates the need for caution in interpreting the results of this test.
TL;DR: It is suggested that at least one of the functions of the alkaline phosphatase of the liver membrane is to hydrolyze phosphorylcholine so that choline can be transported across the bile canalicular membrane.
TL;DR: An alkaline phosphatase was found in sera, ascitic fluid or cancer tissues of patients with hepatocellular carcinoma which had a different electrophoretic mobility from that of liver, bone, placental or intestinal isoenzyme.
TL;DR: Multiple membrane preparations were highly reproducible with respect to the specific activities of the markers studied and an assay suitable for determining 5'-nucleotidase in the small intestine is described.
Abstract: A technique is described for the isolation of a plasma-membrane fraction from the rat intestinal epithelial cell which is distinct from the microvillus membrane of that cell. The isolated fraction contains only about 0.2% of the sucrase activity in the original homogenate and negligible quantities of nuclear and mitochondrial membrane markers. It contains 12% of the total Na+,K+-dependent adenosine triphosphatase and 7% of the alkaline phosphatase, with significant increments in specific activity of these enzymes. Multiple membrane preparations were highly reproducible with respect to the specific activities of the markers studied. The small intestine of one rat yields material containing about 1.3mg of protein. In addition an assay is described suitable for determining 5′-nucleotidase in the small intestine.
TL;DR: Evidence is reported, based on denaturation by heat and on various dietary manipulations in the rat, which suggest that the two enzymes of alkaline phosphatase and Ca ATPase may be part of a single enzyme complex.
Abstract: SEVERAL recent studies suggest that at least part of the effect of vitamin D on intestinal calcium transport may be mediated by a calcium-stimulated ATPase (Ca ATPase) situated in the brush borders of intestinal mucosal cells1. Intestinal brush borders are known to contain several other enzymes which may be involved in digestion and transport, but the relationship between them is poorly understood2. In the case of the disaccharidases, the question of the number of enzymes responsible for the various activities has been raised3. Alkaline phosphatase is also present in brush borders but its function is unknown. During studies of the effect of various agents On intestinal calcium transport we noted a close correlation between the activities of alkaline phosphatase and Ca ATPase. We therefore wondered whether the activities might be due to a single enzyme species. Haussler et al.4 and Norman et al.5 have also noted a close relationship between the two activities under a variety of conditions but Holdsworth6 thinks the two activities are not related. We report here further evidence, based on denaturation by heat and on various dietary manipulations in the rat, which suggest that the two enzymes may be part of a single enzyme complex.
TL;DR: The immunological relationships detected among the alkaline phosphatases of enterobacteria agree approximately with those reported for five other enzymes, as well as with the tryptic peptide pattern similarities found for two other enzymes and with the relationships detected by interspecific deoxyribonucleic acid hybridization tests.
Abstract: An immunological approach has been used for the study of alkaline phosphatase evolution in bacteria of the family Enterobacteriaceae. Antisera were prepared against alkaline phosphatase from Escherichia coli and Klebsiella aerogenes and tested against the unpurified alkaline phosphatases of 32 strains of enterobacteria by double diffusion and quantitative micro-complement fixation. The immunological relationships detected among the alkaline phosphatases of enterobacteria agree approximately with those reported for five other enzymes, as well as with the tryptic peptide pattern similarities found for two other enzymes, and with the relationships detected by interspecific deoxyribonucleic acid hybridization tests.
TL;DR: The temporal relation between tumor removal and apparent cure of VDRR suggests that the tumor was secreting a vitamin D antagonist, and the histologic findings suggested a similar cell type in this case and three similar ones.
Abstract: In a 40-year-old man with bone pain, hypophosphatemia, normocalcemia, elevated serum alkaline phosphatase and increased renal phosphate clearance, and radiologic evidence of osteomalacia, ...
TL;DR: Observations on the incidence of significant titers of neutralizing antibodies to salmon and porcine calcitonin during their chronic (> 4 months) administration to man clearly indicate that an appraisal of this possibility be included in studies involving protracted use of these hormones.
Abstract: 15 patients with Paget's bone disease were treated with varying schedules of porcine (3.8-157.5 MRCU/kg per wk) and/or salmon (1.5-210 MRCU/kg per wk) calcitonins over periods ranging from 4 to 24 months. All of the subjects experienced a striking decrease in serum alkaline phosphatase during the first 4 months of treatment. In six patients, however, resistance to these peptides was suggested by a subsequent elevation of alkaline phosphatase activity in spite of continued and augmented hormone administration. These rebounds in alkaline phosphatase levels correlated with the appearance of calcitonin-binding substances and neutralizing material in serum. Incubations of calcitonins-(125)I and sera from these six subjects resulted in the association of radioactivity with material whose behavior on chromatoelectrophoresis (6/6), sucrose density ultracentrifugation and immunoelectrophoresis (one subject) was identical with that of 7S immunoglobulin. Specific, reversible in vitro binding of salmon calcitonins-(125)I was observed in sera obtained from these patients 5 to 12 months after initiation of salmon calcitonin therapy. All six of these subjects' sera acquired the capacity to neutralize salmon calcitonin's hypocalcemic effect in rat bioassay. Neutralization titers correlated with maximal binding capacities, which ranged from 0.042 to 6.6 mg/liter of serum. Competitive displacement of calcitonins-(125)I from the sera of one patient treated with both porcine and salmon calcitonin indicated separate populations of antibodies to these hormones. In spite of return of disease activity comparable to baseline levels, 3/5 resistant subjects treated with salmon calcitonin failed to develop hypocalcemia after injection of 300-1000 MRCU of salmon calcitonin, but two of these patients developed hypocalcemia in response to the porcine hormone. The disappearance of total radioactivity from the circulation after intravenous administration of salmon calcitonin-(125)I was retarded and the amount of serum radioactivity precipitable in 50% (NH(4))(2)SO(4) greater in 3/3 resistant patients compared to control subjects. These observations on the incidence of significant titers of neutralizing antibodies to salmon (40%) and porcine (66%) calcitonins during their chronic (> 4 months) administration to man clearly indicate that an appraisal of this possibility be included in studies involving protracted use of these hormones.
TL;DR: In general, the largest proteins (including the disaccharidases), were turning over the fastest, and other membrane proteins (i.e. alkaline phosphatase) had an intermediate rate of degradation, and "core" proteins turned over slowly.
Abstract: A B S T R A C T Proteins associated with intestinal brush borders and their various fractions were solubilized with sodium dodecyl sulfate and 8-mercaptoethanol, and separated by electrophoresis on acrylamide gels containing sodium dodecyl sulfate. Brush borders contain at least 15 proteins or subunits, ranging in molecular weight from 19,000 to 270,000. The largest proteins (170-270,000 mol wt), including the disaccharidases, are removed from the brush borders by papain. Proteins belonging to the remaining membrane, including alkaline phosphatase, have an intermediate size (53140,000 mol wt). The proteins corresponding to the filamentous "core" of the microvilli are the smallest (1945,000). The relative rates of degradation of these proteins were studied by following the rate of decline of 14Clabeled leucine activity in specific proteins, and by the double isotope technique of Schimke in which leucine"C was given to intact rats intraluminally 10 hr before an intraluminal dose of leucine-3H. Heterogeneity of 'H/"C ratios and thus of rates of turnover of brush border proteins was noted. In general, the largest proteins (including the disaccharidases), were turning over the fastest. Other membrane proteins (i.e. alkaline phosphatase) had an intermediate rate of degradation, and "core" proteins turned over slowly. Thus, there was a general correlation between relative degradation rate
TL;DR: Polymorphonuclear leukocyte granules were submitted to zonal fractionation through a discontinuous sucrose gradient and enzymatically characterized by peroxidase and alkaline phosphatase activity, respectively.
Abstract: Polymorphonuclear leukocyte granules were submitted to zonal fractionation through a discontinuous sucrose gradient. Azurophilic and specific granules were enzymatically characterized by peroxidase and alkaline phosphatase activity, respectively. The enzymes formed modal distributions like those reported by others. Collagenase activity was consistently associated with the specific granules containing alkaline phosphatase.
TL;DR: Both enzyme activities were very much higher in the Vantress × W.P.R. chicks than in the parent (White Plymouth Rock) strain, but the former birds were less affected by thyrocasein.
TL;DR: The high sensitivity of the electrophoretic method allows the use of normal sera as markers rather than tissue extracts, and isoenzyme patterns may be visually assessed after heat inactivation and chemical inhibition.
Abstract: We describe a new electrophoretic method for the characterization of human serum and tissue alkaline phosphatases on cellulose acetate plates. Enzymes are localized fluorometrically with the substrate α-naphthol AS-MX phosphate or colorimetrically by coupling the reaction product with Fast Blue RR. Both localization techniques are sensitive enough to demonstrate isoenzyme patterns in micro-scale samples of normal sera. Our electrophoretic studies indicate that sera of children and adults normally contain isoenzymes originating from both liver and bone. The high sensitivity of the method allows the use of normal sera as markers rather than tissue extracts, and isoenzyme patterns may be visually assessed after heat inactivation and chemical inhibition. The method is suitable for the electrophoretic fractionation of alkaline phosphatase in large numbers of sera, with equipment and technique familiar to many laboratories.
TL;DR: Experiments with purine-requiring mutants suggest that phosphatase is induced in wild-type strains by an adenine nucleotide, and may be linked to alterations in the levels of the nucleotide pools.
Abstract: Alkaline phosphatase is induced in excess phosphate media by starvation either for pyrimidines or for guanine. Induction is observed both during starvation, after a lag period, and following a period of starvation. Induction is not caused by a lowering of the internal orthophosphate pool, but is linked to alterations in the levels of the nucleotide pools. Experiments with purine-requiring mutants suggest that phosphatase is induced in wild-type strains by an adenine nucleotide. Mutations in the phoR gene can produce differential responses to the different starvation regimes.
TL;DR: In this paper, the epiphyseal plate of the guinea pig was investigated with the electron microscope for the presence of acid phosphatase, ATPase, and alkaline phosphatases activity.
TL;DR: In intestinal fragments from chick embryos and young chicks from 2 days before hatching until 2 days after, the amount of phosphate absorbed from β-glycerophosphate rises in proportion to the increase in phosphatase activity.
TL;DR: The serum isoenzyme level of alkaline phosphatase observed an exponential course as a function of gestation time and the highest level coincided with delivery in half of the patients and in the others, it preceded delivery.
Abstract: By means of a convenient automated method, the placental isoenzyme of alkaline phosphatase in pregnancy serum has been measured in 162 normal pregnancies. The basis of this measurement is, first, the heat inactivation of the nonplacental isoenzyme, and then the measurement of activity (substrate; phenylphosphate) under conditions of pH and substrate concentration which favor the action of the placental isoenzyme. Conditions for a manual method for measuring the placental isoenzyme are also given. The serum isoenzyme level of alkaline phosphatase observed an exponential course as a function of gestation time. The highest level coincided with delivery in half of the patients and in the others, it preceded delivery. On the basis of these normal data, it is now possible to recognize a deviation from the normal curve. The possible significance of the serum placental isoenzyme of alkaline phosphatase is discussed.
TL;DR: The localization of alkaline phosphatase in the enamel organs of 1-day-old rats was investigated using the lead-citrate method and some cytoplasmic bodies seen in the cells of the stratum intermedium occasionally showed enzyme activity.
TL;DR: It is suggested that only the basic form of l-phenylalanine is inhibitory and possible mechanisms are discussed, consistent with a mechanism whereby l- phenylalanines prevents dephosphorylation while other schemes giving rise to uncompetitive inhibition are excluded.
Abstract: The reaction of l-phenylalanine, a stereospecific and uncompetitive inhibitor, with human placental alkaline phosphatase in the free and phosphoryl forms was investigated, using a rapid mixing and quenching technique. It is established that, at pH 8—9, l-phenylalanine reacts instantaneously with phosphoryl phosphatase preventing dephosphorylation at concentrations similar to those causing inhibition in the steady state. It is further established that at pH 9 l-phenylalanine produces an increase in the steady state concentration of phosphoryl phosphatase, amounting to over 80% of the total enzyme with 5 mM l-phenylalanine. At high inhibitor concentration the same steady-state activity was observed with a variety of phosphate ester substrates that different widely in affinity and reactivity. All these results are consistent with a mechanism whereby l-phenylalanine prevents dephosphorylation while other schemes giving rise to uncompetitive inhibition are excluded. It is suggested that only the basic form of l-phenylalanine is inhibitory and possible mechanisms are discussed.
TL;DR: Difference spectroscopy indicated that both of these ligands were bound to the alkaline phosphatase dimer at the same time, related to the catalytic mechanism of this enzyme, with particular reference to the role of two identical subunits in a dimeric enzyme that exhibits only one active site functioning in catalysis at any given time.
Abstract: The temperature-jump technique was used to study the binding equilibrium between the Escherichia coli alkaline phosphatase dimer and 2-hydroxy-5-nitrobenzyl phosphonate in 0.1m-tris buffer, pH8.0. Three partially discrete relaxations were observed, two of which could be related to the bimolecular associations of ligand with different conformations of the enzyme and the third to the interconversion of these states. Relaxation spectra were also used to analyse the changes in the mechanism of ligand binding to alkaline phosphatase caused by increase in ionic strength. The relaxation spectrum observed after the addition of Pi to the equilibrium mixture of phosphonate and enzyme was also studied. Difference spectroscopy indicated that both of these ligands were bound to the alkaline phosphatase dimer at the same time. These results are related to the catalytic mechanism of this enzyme, with particular reference to the role of two identical subunits in a dimeric enzyme that exhibits only one active site functioning in catalysis at any given time.