Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound   Target Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound

---

Query: EC:3.1.3.1 (alkaline phosphatase)
47,916 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

A nucleoside catalyzing the oxidoreduction of NADH and K3Fe(CN)6 was isolated from Torula yeast RNA and also obtained in 0.05% yield by a series of steps: SDS-phenol extraction, nuclease P1 digestion, alkaline phosphatase digestion, anion exchange chromatography, and HPLC on an ODS column. Its chemical structure was clearly determined at 5-hydroxycytidine, from the results of FAB-MS and 1H and 13C NMR spectroscopies. The mass spectra, chromatographic behavior, UV spectra, and NMR spectra of this nucleoside from natural and synthetic sources were identical. This is the first report of an RNA catalyst having catalytic activity except for the cleavage and ligation of phosphodiester bonds of RNA. That an RNA has oxidoreduction activity indicates new possibilities for RNAs as "living molecules". 5-Hydroxycytidine may be a vestige of RNAs that formerly possessed metabolizing ability.
...
PMID:A novel minimum ribozyme with oxidoreduction activity. 227 69

Bulbs of Crocus sativus variety Cartwrightianus were found to contain both a platelet aggregation inducer and inhibitor. The aggregating factor has a Mr of 42 kDa estimated by a Sephadex G75 column and SDS-polyacrylamide slab gel electrophoresis. It was found to lack enzymatic activity such as proteinase, esterase and acid or alkaline phosphatase. The inhibitory factor was also purified to homogeneity by different chromatographic techniques and shows a Mr of 27 kDa as it was estimated by Biogel P30 column and SDS-polyacrylamide slab gel electrophoresis. It was found to possess strong proteinase activity.
...
PMID:Characterization of the platelet aggregation inducer and inhibitor isolated from Crocus sativus. 228 72

When antigens are isolated from staphylococcal protein A immunoprecipitation pellets for analysis by SDS polyacrylamide gel electrophoresis and immunoblotting, severe background problems, due to the presence of antisera and bacterial proteins, can result. We describe a procedure for the analysis of immunoprecipitated systemic lupus erythematosus antigens (e.g., La, Ro, and Sm) which significantly reduces this background while retaining sensitivity with respect to antigen detection. We have adapted a method previously described (MacSween, J.M. and Eastwood, S.L. Methods Enzymol. 1981; 73:459-471) in which lithium diiodosalicylate is used to separate the immunoprecipitated antigen from a covalent antibody-staphylococcal protein A complex. In addition, a modified series of immunoblot incubations was employed, in which antigenic proteins were identified by incubating blots with the antiserum used for the original immunoprecipitation (e.g., La) followed by protein A-biotin and avidin-alkaline phosphatase. Overall, the procedure is straight-forward and may be applicable to other immunoblot systems.
...
PMID:Detection of immunoprecipitated systemic lupus erythematosus antigens by immunoblot analysis. 228 92

Highly purified growth hormone (GH) has been isolated from Atlantic salmon (Salmo salar) pituitaries by extraction with acid acetone, acidic precipitation, and reversed-phase high-performance liquid chromatography (HPLC). The yield was 2.5 mg/g wet tissue. The Atlantic salmon GH (sGH) emerged as a single symmetrical peak after HPLC on a reverse phase C18 column. SDS-gel electrophoresis revealed only one band with an estimated molecular weight of 23,000. Atlantic sGH showed a uniform molecular weight, but two-dimensional (2D) gel electrophoresis of the purified sGH revealed charge heterogeneity with pI's ranging from 6.5 to 8.2. Treatment of the purified sGH with alkaline phosphatase concentrated these different forms into a single more alkaline position (pI 8.2) indicating removal of acidic groups. These results were documented using both silver- and immunostaining of the 2D SDS gels. The purified sGH was phosphorylated in vitro by a calmodulin-dependent protein kinase. Phosphorylation of sGH may be a post-translational modification resulting in several molecular forms with variable acidity. Analysis of the amino acid composition of Atlantic sGH revealed homology with GHs isolated from other teleost species and the amino-terminal sequence showed only three different amino acids within the first 25 residues compared to GH isolated from chum salmon (Oncorhynchus keta) and coho salmon (Oncorhynchus kisutch) pituitaries. Atlantic sGH had a methionine as the amino-terminal residue. Antibodies against chum sGH cross-reacted with Atlantic sGH. Antibodies against either Atlantic or chinook (Oncorhynchus tschawytscha) salmon prolactin or human GH did not cross-react with Atlantic sGH. Atlantic sGH was shown to have a slight growth-promoting activity in the rat tibia assay.
...
PMID:Purification and characterization of Atlantic salmon growth hormone and evidence for charge heterogeneity. 228 75

Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) and quantitative immunoelectrophoretic techniques have been used to characterize further the two major mouse allergens, antigen 1 (Ag 1) and antigen 3 (Ag 3). Gel filtration using Sephacryl S-200 showed Ag 1 to have a molecular weight of 18 kD and Ag 3 of 21 kD. SDS-PAGE followed by Western blotting onto nitrocellulose then incubation with individual antisera directed against each of the two major allergens, and an alkaline phosphatase enzyme system, was used to distinguish between the two allergens and indicated a molecular weight of 17 kD for Ag 1 and 16 kD for Ag 3. Ag 3 but not Ag 1 was shown to contain polysaccharide residues. Immunohistochemical staining of mouse skin sections demonstrated that antigens detected in whole dust extracts were present in the hair follicles, on the hair shafts and on the stratum corneum. Staining of similar sections using the rabbit anti-Ag 3 showed the presence of this major allergen in the hair follicles coating the hairs and extending along the skin surface. Serum from a pool of mouse-allergic subjects also demonstrated staining in the same areas when detected using a fluorescein-labelled anti-human IgE as second antibody. As both major allergens were present in extracts of fur this would appear to be most appropriate for use in diagnosis (i.e. skin test and RAST) and also possibly desensitization. However, dust from isolators (available in greater amounts) would be equally suitable.
...
PMID:Allergy to mice. II. Further characterization of two major mouse allergens (AG 1 and AG 3) and immunohistochemical investigations of their sources. 231 Sep 85

One of two main FL-amnion cell alkaline phosphatase (AP), the fast migrating one (FL-APF) has been reported to be identical to Kasahara isoenzyme (K.I.), which occurs preferentially in sera of patients with primary hepatoma. We purified FL-APF of which the apparent molecular weight was 135,000 by gel filtration, and that of the subunit was 62,000 on SDS/PAGE, indicating homodimeric structure of FL-AL-APF. FL-APF was found to react with monoclonal antibody against adult intestinal AP, but not with monoclonal antibody to placental AP. We isolated FL-APF cDNA clone from FL-amnion cells, of which cDNA was 2525 base pairs in length. Nucleotide sequence of the coding region and the 3' untranslated region was identical to the sequence of human adult intestinal AP cDNA. But the untranslated region of the 5' end of the isolated clone was slightly longer than that of intestinal AP. Hence, FL-APF (K.I.) may occur by altered glycosylation of intestinal AP.
...
PMID:Purification and some properties of the fast migrating alkaline phosphatase in FL-amnion cells (the Kasahara isoenzyme) and its cDNA cloning. 231 Dec 50

1. A heat-stable alkaline phosphatase was purified from Penaeus japonicus, with a final specific activity of 21,280 U/mg of protein. 2. In polyacrylamide-gel electrophoresis under non-denaturing conditions, the purified shrimp alkaline phosphatase was found to have an identical molecular size and surface charge as the human placental enzyme. 3. By using SDS-PAGE, the monomers of shrimp alkaline phosphatase were discovered to have a Mr 55,000 but those of human placental enzyme with a Mr 70,000. Deglycosylation decreases the Mr values of the subunits to 33,000 for shrimp alkaline phosphatase. 4. The purified alkaline phosphatase from shrimp was recovered with both the attachment sites for sialic acids and phosphatidylinositol. 5. The shrimp alkaline phosphatase has an isoelectric point (pI) of 7.6 and the human placental enzyme has a pI of 4.8.
...
PMID:A heat-stable alkaline phosphatase from Penaeus japonicus Bate (Crustacea: Decapoda): a phosphatidylinositol-glycan anchored membrane protein. 233 71

Monoclonal antibodies against the surface of embryonic osteogenic cells have been used to characterize the osteoblastic lineage. One antibody, SB-1, reacts in frozen sections with a family of cells in bone, liver, kidney, and intestine which are identically stained by the histochemical substrate for alkaline phosphatase. In this report, biochemical and immunochemical evidence is presented to indicate that SB-1 is directed against an epitope on alkaline phosphatase which is shared by isoenzymes in a variety of chick tissues. In a solid-phase assay system, high dilutions (1:10(5] of ascites fluid were found to give significant binding of SB-1 to alkaline phosphatase extracted from chick limb or intestine. Partial purification of intestinal alkaline phosphatase on a Sepharose CL-6B column results in the co-elution of alkaline phosphatase enzyme activity and antibody-binding material; this indicates that SB-1 recognizes intestinal alkaline phosphatase rather than an impurity in the crude preparation. Furthermore, Western immunoblots of chick calvarial bone extract electrophoresed on a 5-20% SDS-polyacrylamide gel show that SB-1 reacts with a single 155 kD band which also is stained by the alkaline phosphatase histochemical substrate. In a similar set of experiments, SB-1 reacts with an intestinal alkaline phosphatase isoenzyme whose molecular weight is approximately 185 kD. From these studies, we conclude that SB-1 is specifically reactive with alkaline phosphatase isoenzymes present in bone, liver, kidney, cartilage, and intestine. The reactive epitope is stable to SDS denaturation, not associated with the active site of the enzyme, and dependent on disulfide bonds which impart secondary structure to the protein.
...
PMID:A monoclonal antibody against the surface of osteoblasts recognizes alkaline phosphatase isoenzymes in bone, liver, kidney, and intestine. 235 24

In the midgut tissue of the silkworm, Bombyx mori, alkaline phosphatase isozymes, membrane-bound (m-ALP) and soluble (s-ALP) forms are controlled by non-allelic genes on the same chromosome. We purified and characterized both ALPs to elucidate their possible functions and to compare with mammalian ALPs. Both forms were found to be similar Mr = 68,000 in gel permeation chromatography and as a single subunit as a monomer in SDS-polyacrylamide gel electrophoresis with Mr = 58,000 for m-ALP and Mr = 61,000 for s-ALP. The pH optima of ALPs were 10.9 (m-ALP) and 9.8 (s-ALP), and the former was extremely stable even in pH 10-12 which accords with the physiological milieu in Bombyx midgut lumen. Both ALPs had similar substrate specificities. L-cysteine inhibited strongly both ALPs, but inhibitory effects of L-phenylalanine, L-homoarginine, and L-leucine were undetectable for s-ALP and very weak for m-ALP. The antibody raised against purified m-ALP recognized m-ALP but not purified s-ALP and vice versa. Rocket-immunoassay showed that m-ALP was distributed in similar levels along the length of midgut except for the most anterior portion. Seventy percent of s-ALP activity existed in the last one-third of midgut. Immunohistochemical study revealed that the m-ALP was localized at the brush border of columnar cells in the middle and posterior midgut epithelia. In contrast, the s-ALP was localized at the apical surface of goblet cells through the length of midgut. We detected ATPase activity in the purified s-ALP preparation; Mg2+ was essential for the ATPase activity and the activity also increased with KHCO3 but not with KCl. The solubilization test of m- ALP with various agents was attempted and the relationship between m-ALP and the digestive fluid-ALP was discussed.
...
PMID:Genetically defined membrane-bound and soluble alkaline phosphatases of the silkworm: their discrete localization and properties. 239 71

The chemical nature of association of RNA in immunoprecipitates of human SS-B/La ribonucleoprotein, an autoantigen expressed in various autoimmune disorders, was investigated. A fraction of RNA associated with SS-B/La immunoprecipitates was readily dissociated by SDS-polyacrylamide gel electrophoresis, yielding four main subfractions, R1-4, with chain lengths in the range of 90-130 nucleotides (R4), 140-175 nucleotides (R2 and R3) and above 200 nucleotides (R1). Moreover, the immunoreactive protein component, migrating with a molecular mass of 49 kDa, contained a very tightly bound RNA co-migrating with the protein unless the protein was proteolytically degraded. Most of the RNA molecules in this fraction, represented by about 20 components, had a free 3'-terminus but a blocked 5'-terminus and showed chain lengths between 10 and 125 nucleotides. After pretreatment with alkaline phosphatase and a mixture of ribonucleases T1 + T2 + A, adenosine 3',5'-biphosphate (pAp) was liberated by phosphodiesterase (Crotalus durissus) as the blocked 5'-end of the RNA. The chemical nature of the blockage was revealed after alternative treatment of the protein-pAp component with phosphodiesterase or nuclease S7 followed by acid hydrolysis and phosphoamino acid analysis which showed that a threonine residue must be directly involved in the RNA-protein linkage of 49 kDa SS/La antigen, indicating the presence of a covalent threonine-pAp bond.
...
PMID:Human SS-B/LA autoantigen contains a covalent protein-RNA linkage. 244 50


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

---