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

A conjugate of hippuryllysine (HP) and adenylic acid was synthesized and purified. The structure of the conjugate, hippuryllysyl(N-epsilon-5'-phospho)adenosine (HLAMP) was established using 31P nuclear magnetic resonance, UV spectroscopy, acid/base lability, and enzyme digestion with AMP deaminase, alkaline phosphatase, 5'-nucleotidase, and a phosphoamidase activity recently identified in Dictyostelium discoideum. The results indicate that HLAMP contains a phosphoamide bond between the phosphate of AMP and the epsilon amino group of HL. Employing a microdroplet assay to assess chemotactic activity, HLAMP was found to be a potent chemoattractant of 7-h developing amoebae of D. discoideum. Other conjugates, including lysine-AMP (LAMP), tuftsin-AMP (TAMP) and avidin-AMP (AVAMP), as well as the degradation products of HLAMP (HL, AMP, and lysine) exhibited no chemotactic activity. The molecular structure of HLAMP is compared to that of other known chemoattractants of the cellular slime molds, and possible chemotactic receptors for HLAMP are considered.
 ...
PMID:HLAMP--a conjugate of hippuryllysine and AMP which contains a phosphoamide bond--stimulates chemotaxis in Dictyostelium discoideum. 344 32

The degree of phosphorylation of intestinal microvilli membrane proteins in an adult amphibian, Rana esculenta, was investigated under various experimental conditions. The microvilli protein phosphorylation rate rapidly increases during the first 4 min of incubation in a medium containing [gamma-32P]ATP. This increase is slower afterwards. Cyclic nucleotides (cyclic AMP, cyclic GMP) and sorbitol do not modify the microvilli protein phosphorylation rate. On the contrary, this phosphorylation rate significantly decreases in the presence of L-lysine, when its concentration in the incubation medium is greater than 25 mM. The time course of phosphorylation confirms the inhibitory effects of L-lysine (100 mM). The microvilli membrane proteins were distinguished by polyacrylamide gel electrophoresis. In heated samples, electrophoresis followed by an radioautograph systematically reveals the existence of a very phosphorylated protein with a mol. wt of 86 kDa. The phosphorylation of this protein is partially inhibited by L-lysine (100 mM). The very phosphorylated protein could be the monomer of alkaline phosphatase. The dimer (170 kDa) is visualized on electrophoretograms by its catalytic activity. In mammals, several authors have established a correlation between phosphorylation of the microvilli membrane proteins and the intensity of intestinal calcium absorption. Such a control is presently being investigated in adult Rana esculenta.
 ...
PMID:Phosphorylated proteins from anuran intestinal microvilli membranes--I. Relations with alkaline phosphatase. 348 10

Fluorescein (Fl) and tetramethyl rhodamine (Rh) were evaluated as possible candidates for a double hapten sandwich system in enzyme immunohistology. Monoclonal antibodies were raised against Fl and Rh. Their fine-specificity was tested with a competition-like assay. A pair of Mab's was selected for immunohistology in which they functioned as a bridge between Fl/Rh conjugated antibodies and Fl/Rh labeled peroxidase and alkaline phosphatase, respectively. The binding of fluorescein labeled antibodies could be successfully demonstrated in histological slides. A large variability in the efficacy of staining was observed in the case of rhodamine labeled antibodies. The phenomenon is explained by assuming that tetramethyl rhodamine isothiocyanate reacts preferentially with lysine residues near to, or embedded in, hydrophobic regions in a protein. This condition may reduce the accessibility of the Rh moiety for anti-Rh antibodies.
 ...
PMID:Fluorescein and tetramethyl rhodamine as haptens in enzyme immunohistochemistry. 352 96

Monoclonal antibodies (McAb) were used to develop nonisotopic and radioimmunoassays (RIA) for quantitative determination of the major nicotine metabolite, cotinine, in physiological fluids. ELISAs and fluorescence immunoassays were carried out in microtiter plate wells coated with a conjugate of cotinine 4'-carboxylic acid bound covalently to poly-L-lysine. The detection systems were horseradish peroxidase (HRP)-labeled staphylococcal protein A, HRP-streptavidin-biotin, and biotinylated alkaline phosphatase-4-methylumbelliferyl phosphate. With the three McAb tested, I50 values ranged between 0.024-0.063 ng cotinine and as little as 0.005-0.015 ng gave 15% inhibition. These assays were 5-20 times more sensitive than similar assays using six rabbit antisera. With McAb the standard inhibition curves were steeper and complete inhibition of immune binding was achieved with approximately 1 ng cotinine. In contrast, 100-500 ng cotinine failed to give greater than 80-90% inhibition with rabbit antibodies either in the plate assays or in RIA using a 125I-labeled tyramine derivative of cotinine as the tracer. In this RIA, the sensitivity with McAb (mean I50 of 0.55 ng cotinine) was over three-fold greater than with rabbit antisera (mean I50 of 1.84 ng). The presence of antibodies directed to the amide linkage group common to the polylysine conjugate. 125I-tyramine derivative and the immunogen likely accounts for the inferior quality of assays using rabbit antisera. Consistent with this conclusion, superimposable inhibition curves were obtained in the RIA when monoclonal or rabbit antibodies were used with [3H]cotinine. Cotinine levels in saliva, serum and plasma from smokers and non-smokers determined with McAb-based assays showed a strong correlation with values obtained by RIA using rabbit antisera or by gas chromatography. Properly selected McAb offer distinct advantages over conventional antisera in nonisotopic immunoassays and RIAs for cotinine as a biochemical marker of active or passive smoking.
 ...
PMID:Comparison of monoclonal and polyclonal antibodies to cotinine in nonisotopic and isotopic immunoassays. 354 36

Experiments were conducted to examine the interrelationships between methionine, choline and inorganic sulfate in the diet of weanling pigs, and to evaluate the selenium (Se) status of pigs fed diets with or without supplemental sulfate. Two trials utilized 288 weanling (3-wk-old) pigs allotted to dietary treatment based on weight, sex and litter origin. There were six pigs/pen and three replicate pens/treatment in each trial. The basal corn-soybean meal diet was formulated to supply .55% sulfur amino acids and contained a choline and sulfur-free vitamin and mineral premix. Lysine was added to provide a total of 1.13% lysine. Seven additional treatments were formulated by substituting for corn .17% DL-methionine, .29% choline dihydrogen citrate or .25% Na2SO4 to create a 2(3) factorial arrangement of treatments. There were methionine X choline X sulfate interactions for average daily gain (P less than .001) and feed-to-gain ratio (F:G; P less than .05). Adding choline, methionine, Na2SO4 or choline plus methionine to the basal diet did not improve gains. However, when Na2SO4 plus methionine or Na2SO4 plus choline were added, daily gains were increased (P less than .05) and F:G was improved (P less than .1). Addition of all three supplements did not result in a further increase in gain. Pigs fed choline-supplemented diets had higher (P less than .01) hematocrit and tended (P = .07) to have increased hemoglobin concentration. There was no effect on serum triglycerides or alkaline phosphatase activity due to dietary treatment. The concentration of Se in muscle, liver, kidney and blood was not influenced by sulfate content of the diet.
 ...
PMID:Methionine, choline and sulfate interrelationships in the diet of weanling swine. 375 83

An enzyme-linked immunosorbent assay (ELISA) using plates coated with hepatocyte plasma membranes (HPM) was developed for the measurement of antibodies directed at hepatocyte surface antigens. Precoating ELISA plates with poly-L-lysine (PLL) provided firm attachment for the adsorption of HPM. The use of HPM, in preference to whole hepatocytes, excludes pathologically irrelevant cytoplasmic antigens. In addition, there is no necessity for glutaraldehyde fixation which is commonly used in cellular assays to maintain cellular integrity and which may result in loss or alteration in antigenic specificities. The assay was used to study loss of tolerance to mouse HPM in mice immunized with rat HPM. Three mouse strains were immunized, each strain developed antibodies to rat HPM and autoantibodies to mouse HPM with autoantibody levels reaching a peak 6-10 weeks after commencement of immunization. The correlation between ELISA and indirect immunofluorescence for the measurement of HPM autoantibodies was 0.79 (P less than 0.001) within the serum titration range of 1:25 to 1:200. Antibody to control kidney plasma membrane (KPM) was also measured by ELISA, after elimination of endogenous alkaline phosphatase activity using levamisole. Immunization with rat HPM elicited organ-non-specific autoantibodies to KPM, but these were at lower levels than autoantibodies to HPM.
 ...
PMID:An enzyme-linked immunosorbent assay for the detection of hepatocyte plasma membrane antibodies. 390 61

Young rats were force-fed a lysine + arginine-devoid diet or a complete diet for 3 days, and selected biochemical and morphologic studies were conducted. Rats force-fed the experimental diet in comparison with those force-fed the control diet for 3 days showed decreased body weight gain, hepatomegaly with periportal fatty liver, pancreatic and splenic atrophy, and enhanced 14C-leucine incorporation into hepatic proteins. Differences in the experimental animals were observed in the free amino acid levels of serum (decreased lysine, arginine, and ornithine) and liver (decreased ornithine), in blood chemistries (decreased levels of ammonia N2, uric acid, cholesterol, protein, albumin, alkaline phosphatase, LDH and SGOT) and in hematologic findings (leukocytopenia and thrombocytopenia after a morning feeding). The experimental findings in young rats force-fed the lysine + arginine-devoid diet were compared with those reported to develop in children with lysinuric protein intolerance (LPI), an autosomal recessive defect in diamino acid transport. Children with LPI as described by others reveal a number of similarities as well as a number of differences in comparison to the findings in the experimental animals. The comparison suggests that some of the pathological manifestations of LPI may be related to a deficiency of diamino acids but others must be due to different alterations in this complex human disease.
 ...
PMID:Chemical pathology of diamino acid deficiency: considerations in relation to lysinuric protein intolerance. 393 96

Binding of fibronectins to gangliosides was tested directly using several different in vitro models. Using an enzyme-linked immunoabsorbent assay (ELISA), gangliosides were immobilized on polystyrene tubes and relative binding of fibronectin was estimated by alkaline phosphatase activity of conjugated second antibody. Above a critical ganglioside concentration, the gangliosides bound the fibronectin (GT1b congruent to GD1b congruent to GD1a greater than GM1 much greater than GM2 congruent to GD3 congruent to GM3) in approximately the same order of efficiency as they competed for the cellular sites of fibronectin binding in cell attachment assays (Kleinman et al., Proc natl acad sci US 76 (1979) 3367). Alternatively, these same gangliosides bound to immobilized fibronectin. Rat erythrocytes coated with gangliosides GM1, GD1a or GT1b bound more fibronectin than erythrocytes not supplemented with gangliosides. Using fibronectin in which lysine residues were radioiodinated, an apparent Kd for binding to mixed rat liver gangliosides of 7.8 X 10(-9) M was determined. This value compared favorably with the apparent Kd for attachment of fibronectin to isolated plasma membranes from rat liver of 3.7 X 10(-9) M for fibronectin modified on the tyrosine residue, or 6.4 X 10(-9) M for fibronectin modified on lysine residues. As shown previously by Grinnell & Minter (Biochem biophys acta 550 (1979) 92), fibronectin modified on tyrosine residues did not promote spreading and attachment of CHO cells. It did, however, bind to cells. In contrast, lysine-modified fibronectin both bound to cells and promoted cell attachment. Plasma membranes isolated from hepatic tumors in which the higher gangliosides that bind fibronectin were depleted bound 43-75% less [125I]fibronectin than did plasma membranes from control livers. The findings were consistent with binding of fibronectins to gangliosides, including the same gangliosides depleted from cell surfaces during tumorigenesis in the rat.
 ...
PMID:Fibronectin binding to gangliosides and rat liver plasma membranes. 394 47

Intake of the essential amino acids, threonine, lysine and methionine by Ukrainian children of different height was studied. The processes of osteogenesis and phosphorus-calcium metabolism in rats with the above amino acids deficiency in the diet were also subjected to study. A direct correlation was established between intake of the amino acids under study and the height of schoolchildren. The deficiency of the amino acids in the diet of experimental animals contributed to the retardation of the growth, destructive changes, an increase in the content of hydroxyproline, a reduction of phosphomonoesterase-I activity in bones, and alterations in phosphorus-calcium metabolism.
 ...
PMID:[Effect of amino acid deficiency on bone tissue growth and formation]. 403 75

A 125-kilodalton (kDa) phosphoprotein was isolated from nucleoli of Novikoff hepatoma cells in the presence of various inhibitors of proteases, alkaline phosphatase, and RNase. This protein was the most highly phosphorylated protein found thus far in the nucleolus. The half-life of [32P]phosphate in the 125-kDa phosphoprotein was approximately 60 min. Amino acid analysis of the protein showed it had a high serine content (15.5 mol %), a high glutamine plus glutamic acid content (15.5 mol %), and a high lysine content (10.3 mol %). Phosphoserine was the only phosphorylated amino acid identified. After alkaline hydrolysis of the 32P-labeled protein, ribonucleotides were found which accounted for approximately 8.5% of the [32P]phosphate. After cytidine 3',5'-[32P]diphosphate ([32P]pCp) labeling by RNA ligase, several oligoribonucleotide sequences were purified including GGGCOH and GGGGCOH. The binding of oligonucleotides to peptides was stable under denaturing fractionation conditions including 6 M urea treatment and incubation at 100 degrees C for 10 min in sodium dodecyl sulfate and beta-mercaptoethanol. Furthermore, when nucleotide-peptide complex was treated with ribonuclease T2 followed by snake venom phosphodiesterase, the junctional nucleotide pCp was released. These results suggest that one or more ribonucleotides are covalently bound to the 125-kDa phosphoprotein.
 ...
PMID:Isolation and characterization of a 125-kilodalton rapidly labeled nucleolar phosphoprotein. 408 83


<< Previous 1 2 3 4 5 6 7 8 9 10 Next >>