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)

Two matched groups of postmenopausal patients were treated respectively with calcitonin or calcitonin and an arginine-lysine-glycerophosphoric acid-lactose association. The rationale underlying this therapy took the form of data in the literature which indicated an action of these amino acids and lactose on calcium absorption and on the metabolism of protein components in the skeletal structure. The following tests were performed: mineralometric evaluation, evaluation of painful symptoms and intake of pain-relieving drugs, serum levels of calcium, phosphorus, alkaline phosphatase, osteocalcin, parathormone, and calciuria and hydroxyproline. These parameters were assayed at the beginning and end of treatment which lasted six months. The results, or in other words the comparison between the two groups, basal or after treatment, and the values recorded before and after treatment in each group, enable the authors to affirm that the administration of the arginine-lysine-glycerophosphoric acid-lactose association leads to an increase in bone density and plasma osteocalcin, a reduction in painful symptoms and analgesic intake, and a reduction in the serum levels of parathromone and hydroxyproline. Data reported in the literature support the conclusion that the results obtained are the consequence of an improved intestinal absorption calcium. It is highly probable that the protein components of the association administered, arginine-lysine-glycerophosphoric acid-lactose, also exercise a direct action on osteoblasts and on the metabolism of bone matrix protein components.
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PMID:[Experience regarding the use of arginine-lysine-lactose treatment in menopausal osteoporosis]. 808 36

FtsH is an Escherichia coli protein with its amino-terminal region anchored to the cytoplasmic membrane and with its cytoplasmic domain significantly homologous to the members of an ATPase family found in eukaryotic cells. We previously showed that the loss of ftsH function results in reduced cytoplasmic retention of the alkaline phosphatase moiety that was attached to cytoplasmic regions of membrane proteins (the Std phenotype) and also in translocation retardation of some exported proteins. We now report that expression of some FtsH variants from plasmids causes dominant Std and translocation phenotypes. Such variants include carboxyl-terminally truncated forms of FtsH and some missense mutations in the cytoplasmic domain, notably Lys to Asn changes at the two ATP binding consensus sequences. In contrast, amino-terminally truncated variants lacked the dominant phenotypes. It was suggested that the amino-terminal membrane region of FtsH interacts with other component(s), and that the two putative ATP binding sites are of vital importance for the FtsH functions.
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PMID:Involvement of FtsH in protein assembly into and through the membrane. II. Dominant mutations affecting FtsH functions. 810 5

Ligand-mediated approaches to gene transfer offer an alternative to viral vectors for both in vivo and in vitro applications. Although a significant percentage of the plasmid-based DNA complex is lost to lysosomal degradation following receptor-mediated endocytosis, simultaneous infection with adenovirus has been shown to increase the level of transgene expression [Curiel, Agarwal, Wagner and Cotten (1991) Proc. Natl. Acad. Sci. U.S.A. 88, 8850-8854; Wagner, Zatloukal, Cotten, Kirlappos, Mechtler, Curiel and Birnstiel (1992) Proc. Natl. Acad. Sci. U.S.A. 89, 6099-6103]. In this study we describe an adenovirus-based ligand complex where the plasmid DNA, polycation-ligand conjugate and adenovirus are contained within a single particle structure. At the core of the transfection particle is a replication-defective recombinant adenovirus encoding a cDNA minigene for human placenta alkaline phosphatase that was chemically modified with poly(L-lysine) (Ad-pLys). Electron microscopy of an adenovirus-based ligand complex formed by successively adding plasmid DNA and an asialo-orosomucoid-poly(L-lysine) conjugate to Ad-pLys revealed structures that appeared as intact viral particles coated with a dense biomolecular layer. Adenovirus-based ligand complexes containing either a luciferase or beta-galactosidase reporter plasmid were shown to efficiently deliver the plasmid transgene to cells that express the hepatic asialoglycoprotein receptor. Furthermore, the poly(L-lysine) modification greatly reduced the infectivity potential of the virus without causing a concomitant loss of augmented gene transfer. As an alternative to infectious virions, incomplete products of viral assembly were also considered as a source for endosomalytic activity. However, these defective virions were unable to significantly enhance plasmid transgene delivery.
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PMID:Biochemical and functional analysis of an adenovirus-based ligand complex for gene transfer. 816 59

Ovomucoids are commonly present in bird egg white and exhibit inhibitory activity toward various serine proteases. To investigate the structure-function relationship of ovomucoid domain 3, we established a secretory expression system for the chicken ovomucoid domain 3 (OMCHI3)-encoding gene in Escherichia coli by ligating it downstream from the tac promoter and signal peptide of E. coli alkaline phosphatase. E. coli JM105 was transformed with the resulting plasmid and induced with 1 mM isopropyl-beta-D-thiogalactopyranoside (IPTG). The mature OMCHI3 was detected in the culture supernatant, and was purified to homogeneity by three-step chromatography. Amino-acid sequence analysis showed that processing by the signal peptidase was carried out exactly at the expected site. Measurements of circular dichroism spectra and inhibitory activity indicated that OMCHI3 was produced in the properly folded form. Furthermore, site-specific replacement of the Ala residue at the P1 site with Met or Lys resulted in acquisition of inhibitory activity toward chymotrypsin or trypsin, respectively, indicating that the P1 site is the predominant determinant for inhibitory specificity.
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PMID:Secretory production of chicken ovomucoid domain 3 by Escherichia coli and alteration of inhibitory specificity toward proteases by substitution of the P1 site residue. 820 80

The interactions of bone cells with their surrounding extracellular microenvironment may be mediated by integrins, a family of heterodimeric glycoproteins consisting of alpha and beta subunits that noncovalently interact to form cell-substratum adhesion receptors. We previously described the integrins on calvarial bone cells in rats with use of polyclonal antibodies against some integrin subunits. In the present study, we expanded this initial characterization by employing a more complete panel of monoclonal antibodies to identify integrins on human bone cells. Minced fragments of trabecular bone obtained during total knee arthroplasty were grown in culture until bone cells became confluent. The cells then were dissociated, plated again, grown to confluence, and assayed for alkaline phosphatase activity, response of cyclic adenosine monophosphate to stimulation with parathyroid hormone, and osteocalcin content. The percentage of the cells that adhered to various substrates was measured; 60-70% adhered to type-I collagen, fibronectin, vitronectin, and poly-D-lysine; 40-50% adhered to type-IV collagen, laminin, and gelatin; and only 10% adhered to fibrinogen. Flow cytometric analysis with anti-integrin monoclonal antibodies and sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of immunoprecipitates of the human bone cells revealed high levels of alpha 1 beta 1, alpha 3 beta 1, alpha 5, beta 1 and alpha v beta 5 integrins and much lower levels of alpha 2 beta 1, alpha 4 beta 1, alpha v beta 1, and alpha v beta 3 integrins. This description of the integrin repertoire of cultured human bone cells represents the first step toward an understanding of the role played by integrins in the growth, maintenance, and repair of bone.
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PMID:Identification of integrin receptors on cultured human bone cells. 820 92

Chromogranin A is a secretory protein expressed widely in neuroendocrine cells. It is known to be phosphorylated but the precise sites of phosphorylation are not known. We have isolated, from bovine pancreas and ileum, chromogranin A fragments corresponding to a region giving rise to a biologically active product, pancreastatin. Phosphorylation patterns were determined by fast atom bombardment mass spectrometry and alkaline phosphatase digestion followed by ion-exchange chromatography and radioimmunoassay. In the pancreas, there were unmodified, mono- and di-phosphorylated forms of the fragment chromogranin A(248-313) with Arg and Glu at positions 293 and 301 respectively; in addition, there were small amounts of monophosphorylated peptide with an alternative primary sequence of His and Lys at 293 and 301 respectively. Two products of cleavage, pancreastatin and the fragment 297-313, were also found in unmodified and monophosphorylated forms. In the ileum, peptides with both alternative primary sequences were found, pancreastatin was absent, and phosphorylation was generally less than in the pancreas. Chromogranin A-derived peptides therefore exhibit tissue-specific patterns of phosphorylation and cleavage, and at least two phosphorylation sites occur in the region giving rise to a biologically active product.
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PMID:Post-translational processing of chromogranin A: differential distribution of phosphorylated variants of pancreastatin and fragments 248-313 and 297-313 in bovine pancreas and ileum. 824 Feb 72

Site-specific mutagenesis was used to explore the roles of the side chains of residues Lys-328 and Asp-153 in Escherichia coli alkaline phosphatase. The D153H enzyme exhibits a 3.5-fold decrease in activity at pH 8.0 compared to that of the wild-type enzyme, while a double mutant D153H/K328H exhibits a 16-fold decrease in activity under these conditions. However, the Km values for both enzymes, employing the substrate p-nitrophenyl phosphate, are lower than the value for the wild-type enzyme. The Ki for phosphate, which is pH- and Mg(2+)-dependent, is decreased for the D153H enzyme and increased for the D153H/K328H enzyme. Relative to the wild-type enzyme, both mutant enzymes bind Mg2+ more weakly and undergo a time-dependent activation induced by Mg2+. The half-time of the activation process is independent of the Mg2+ concentration, indicating that the activation most probably involves a conformational change. The pH versus activity profiles of both enzymes are altered relative to that of the wild-type enzyme and exhibit greatly enhanced activity, relative to that of the wild-type enzyme, at high pH values. The pre-steady-state kinetics for the D153H and D153H/K328H enzymes exhibit a transient burst of product formation at pH 8.0, under conditions at which the wild-type enzyme exhibits no transient burst, indicating that at pH 8.0 the hydrolysis of the covalent enzyme-phosphate complex is rate-determining and not the release of phosphate from the noncovalent enzyme-phosphate complex as is observed for the wild-type enzyme. Therefore, these mutations are directly influencing catalysis.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:Magnesium in the active site of Escherichia coli alkaline phosphatase is important for both structural stabilization and catalysis. 843 39

The arcDABC operon of Pseudomonas aeruginosa encodes the enzymes of the arginine deiminase pathway and is induced by oxygen limitation. The arcD gene specifies a 53-kDa protein with arginine: ornithine exchange activity. The ArcD protein of P. aeruginosa, like the LysI lysine transporter of Corynebacterium glutamicum, has 13 hydrophobic regions which could span the cytoplasmic membrane. Fusion of a Caa (colicin A) epitope to the N-terminal part of ArcD permitted the localization, by immunoblotting, of the hybrid protein in the inner membrane of P. aeruginosa. Fusion of PhoA (alkaline phosphatase) to the very C terminus of ArcD produced another hybrid protein, which exhibited PhoA activity. Both ArcD hybrid proteins retained arginine transport activity and served to support a topological model which proposes that the N terminus is oriented toward the cytoplasm and the C terminus faces the periplasm. Further ArcD-PhoA fusions were consistent with this model. When the Caa epitope was fused to a C-terminal ArcD fragment consisting of only 5 hydrophobic domains, the resulting hybrid protein could be recovered intact from the inner membrane, suggesting that the C-terminal part of ArcD contains sufficient information for insertion into the membrane. This study illustrates the utility of the Caa epitope to tag membrane proteins.
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PMID:Characterization of the arcD arginine:ornithine exchanger of Pseudomonas aeruginosa. Localization in the cytoplasmic membrane and a topological model. 844 2

Prothymosin alpha is post-translationally modified. When human myeloma cells were metabolically labeled with [32P]orthophosphoric acid, they synthesized [32P]prothymosin alpha. The incorporated radioactivity was resistant to DNase and RNases A, T1, and T2, but could be completely removed by alkaline phosphatase. No evidence was found for an RNA adduct as postulated by Vartapetian et al. [Vartapetian, A., Makarova, T., Koonin, E. V., Agol, V. I., & Bogdanov, A. (1988) FEBS Lett. 232, 35-38]. Thin-layer electrophoresis of partially hydrolyzed [32P]prothymosin alpha indicated that serine residues were phosphorylated. Analysis of peptides derived from bovine prothymosin alpha and human [32P]prothymosin alpha by treatment with endoproteinase Lys-C revealed that the amino-terminal 14-mer, with serine residues at positions 1, 8, and 9, was phosphorylated at a single position. Approximately 2% of the peptide in each case contained phosphate. Further digestion of the phosphopeptide with Asp-N followed by C18 reversed-phase column chromatography produced two peptides: a phosphate-free 9-mer containing amino acids 6-14 and a labeled peptide migrating slightly faster than the N-terminal 5-mer derived from the unmodified 14-mer. Positive identification of the phosphorylated amino acid was obtained by colliding the 14-residue phosphopeptide with helium in the mass spectrometer and finding phosphate only in a nested set of phosphorylated fragments composed of the first three, four, and five amino acids. The results prove that prothymosin alpha contains N-terminal acetylserine phosphate. In a synchronized population of human myeloma cells, phosphorylation occurred throughout the cell cycle. Furthermore, prothymosin alpha appeared to be stable, with a half-life slightly shorter than the generation time. Although prothymosin alpha is known to be essential for cell division, the constancy of both the amount of the protein and the degree of its phosphorylation suggests that prothymosin alpha does not directly govern mitosis.
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PMID:Phosphorylation of human and bovine prothymosin alpha in vivo. 848 35

The ovarian fluid of rainbow trout (Oncorhynchus mykiss), charr (Salvelinus alpinus), lake trout (Salmo trutta f lacustris) and Danube salmon (Hucho hucho) was analyzed for its inorganic and organic composition. The qualitative composition of the ovarian fluid of the investigated species was similar, but significant quantitative differences were found. The following components were determined: sodium 106.6-142.2 mmol/l, potassium 1.7-2.7 mmol/l, calcium 0.45-0.61 mmol/l, osmolality 256.4-291.6 mosmol/kg, pH 8.4-8.8, glucose 1,698-4,195 mumol/l, fructose 17-399 mumol/l, lactate 34-227 mumol/l, cholesterol 650-1,230 mumol/l, phosphatidylcholine 0.25-3.0 mumol/l, lysophosphatidylcholine 10-100 mumol/l, choline 0-1.1 mumol/l, protein 95.0-278.4 mg/100 ml. Arginine, cystine, glycine, histidine, lysine, proline, serine, tyrosine and valine were the free amino acids occurring in concentrations of more than 10 mumol/l. Activities of alkaline phosphatase (200-6,000 mumol substrate/l/h), lactate dehydrogenase (9-690 mumol substrate/l/h), beta-D-glucuronidase (70-410 mumol substrate/l/h), proteases (140-215 mumol/l/h with collagen substrate, 25-90 mumol/h/l with gelatine substrate) and acid phosphatase (100-130 mumol substrate/l/h) were measured, but not the glucose-6-phosphate dehydrogenase and alpha-glucosidases activities.
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PMID:Composition of the ovarian fluid in 4 salmonid species: Oncorhynchus mykiss, Salmo trutta f lacustris, Salvelinus alpinus and Hucho hucho. 852 77


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