Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:3.5.1.4 (deaminase)
 5,113 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Portions of closed jejunal biopsies from the dog were homogenised and their organelles separated by isopycnic centrifugation on continuous sucrose density gradients. The distributions of marker enzymes for the principal organelles were determined using highly sensitive assay procedures. The following organelles, with assayed marker enzymes and modal densities between brackets were characterised: peroxisomes (catalase, 1.21); brush borders (zinc-resistant alpha-glucosidase, leucyl-beta-naphthyl-amidase, gamma-glutamyl transferase, alkaline phosphatase, 1.20); lysosomes (N-acetyl-beta-glucosaminidase, alpha-mannosidase, 1.19); mitochondria (malate dehydrogenase, 1.18); endoplasmic reticulum (Tris-resistant alpha-glucosidase, 1.16); basal-lateral membranes (5'-nucleotidase, 1.11) and cytosol (lactate dehydrogenase). Homogenisation in isotonic sucrose containing digitonin (0.12 mmol/litre) selectively disrupted lysosomes and increased the equilibrium density of brush border and basal-lateral membranes. This procedure will be used to study the subcellular pathology of naturally occurring intestinal disease in the dog.
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PMID:Subcellular fractionation studies on peroral jejunal biopsies from the dog. 3 Jan 25

Cells from stationary-phase cultures of two strains of Saccharomyces cerevisiae (3 and 20) failed to flocculate when grown in a complex or a chemically defined medium, while those of two other strains (11 and 13) flocculated when grown in either medium. Strain 30 flocculated when grown in complex but not defined medium and harvested from stationary-phase cultures. pH-electrophoretic mobility measurements on all five strains showed that mobility attributable to carboxyl groups usually increased as cultures progressed from the exponential to the stationary phase, while that caused by phosphate groups tended to decline. Acquisition of flocculating ability was accompanied in strains 11 and 30 by a slight increase in amidase activity, and greater increases compared with nonflocculent populations in activities of leucine aminopeptidase. alpha-mannosidase, and proteinase C. Activities of proteinases A and B showed no correlation with acquisition of flocculating ability.
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PMID:Changes in electrophoretic mobility and lytic enzyme activity associated with development of flocculating ability in Saccharomyces cerevisiae. 39 72

The glycoprotein nature of the binding subunit of the dopamine D2 receptor in rat striatum has been examined by photoaffinity labeling receptor preparations with N-(p-azido-m-[125I]iodophenethyl)spiperone followed by treatment of crude membrane receptor or receptor fractions isolated from sodium dodecyl sulfate (SDS) polyacrylamide gels with endo- and exoglycosidases. The major photoaffinity labeled protein migrates as a heterogeneous species on 10% SDS polyacrylamide gels and ranges from 130,000 to 75,000 relative molecular mass (Mr). This heterogeneity can be explained by glycosylation of the receptor by complex-type N-linked oligosaccharides. Three fractions of labeled receptor were isolated from SDS polyacrylamide gels over a range of 130,000 to 75,000 Mr; after digestion with peptide-N4-[N-acetyl-beta-glucosaminyl] asparagine amidase, all fractions yielded a single peptide approximately 40,000 Mr. Treatment of photoaffinity labeled membranes with alpha-mannosidase was without effect. The dopamine D2 receptor appears to contain substantial amounts of sialic acid as treatment of photoaffinity labeled membranes with neuraminidase increased the receptor mobility on SDS polyacrylamide gels to a species of 50,000-54,000 Mr. Treatment of the receptor with neuraminidase followed by endo-alpha-N-acetylgalactosaminidase did not change the electrophoretic migration pattern from that seen after neuraminidase treatment alone, suggesting that the binding peptide contains no serine- or threonine-linked oligosaccharides. A smaller binding peptide of approximately 31,000 Mr is also apparent in crude photoaffinity labeled membranes. This material also contains N-linked oligosaccharide. Complete removal of N-linked oligosaccharide from the dopamine D2 receptor did not change the rank order potency of agonist and antagonist compounds to compete for [3H]spiperone binding to crude membrane fractions. The dopamine D2 receptor represents a highly glycosylated neural receptor.
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PMID:N-linked oligosaccharides are responsible for rat striatal dopamine D2 receptor heterogeneity. 252 26

The binding subunit of the alpha 1-adrenergic receptor has been identified as an Mr = 80,000 peptide in several tissues. Adsorption of the alpha 1-adrenergic receptor to a wheat germ agglutinin lectin-agarose resin suggests that the receptor protein is glycosylated. In this study, we investigated the nature of the carbohydrate chains linked to the alpha 1-adrenergic receptor peptide. The alpha 1-adrenergic receptor from DDT2 MF-2 smooth muscle cell and rat brain membranes was photolabeled with 125I-azido-prazosin [( 125I]CP65,526) and then treated with exoglycohydrolases prior to SDS-PAGE and autoradiography. Removal of terminal sialic acid residues by neuraminidase decreased the receptor Mr by 6,000; however, alpha-mannosidase was without effect, indicating complex type glycosylation of the receptor-protein. Similar results were observed for the rat hepatic membrane alpha 1-adrenergic receptor. Removal of N-linked carbohydrates at asparagine residues by peptide-N4[N-acetyl-beta-glucosaminyl]asparagine amidase (from Flavobacterium meningosepticum) resulted in a specifically labeled peptide at Mr = 50,000-55,000 in DDT1 MF-2 membrane and solubilized receptor preparations. Treatment of DDT1 MF-2 cells with swainsonine or (+)-1-deoxymannojirimycin, inhibitors of complex type carbohydrate chain biosynthesis, caused a reduction in the apparent molecular weight of the receptor (Mr = 60,000) but did not alter the number of alpha 1-adrenergic receptors per cell or their affinity for the radioligand [3H]prazosin. These findings indicate that the alpha 1-adrenergic receptor is heavily glycosylated, the major oligosaccharide moiety being of the complex type, N-linked to asparagine residues. The peptide backbone of the receptor has an Mr less than or equal to 55,000, consistent with the predicted molecular mass of other membrane neurotransmitter receptors based on sequence analysis of isolated cDNA clones.
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PMID:Glycosylation of the mammalian alpha 1-adrenergic receptor by complex type N-linked oligosaccharides. 282 78

1. Ovalbumin glycopeptides, freed from all amino acids other than aspartic acid and a small proportion of leucine by repeated digestion with Pronase, were hydrolysed by 1-aspartamido-beta-N-acetylglucosamine amidohydrolase (glycoaspartamidase) to the corresponding oligosaccharides. The glycoaspartamidase did not attack ovalbumin itself. 2. Ovalbumin, with mannose/hexosamine ratio 5:4, lost 1.5moles of N-acetylglucosamine and more than 2moles of mannose after incubation with alpha-mannosidase and beta-N-acetylglucosaminidase respectively. 3. In ovalbumin glycopeptides with approximate mannose/hexosamine ratios 5:3 and 5:4, one and two N-acetylglucosamine residues respectively were accessible to the action of beta-N-acetylglucosaminidase. 4. A mixture of alpha-mannosidase and beta-N-acetylglucosaminidase, acting on an ovalbumin glycopeptide with mannose/hexosamine ratio 5:3.7, removed nearly 4moles of mannose and 1.5moles of N-acetylglucosamine. 5. alpha-Mannosidase removed about 1.5moles of mannose from the ovalbumin oligosaccharide with mannose/hexosamine ratio approx. 5:3. The subsequent action of beta-N-acetylglucosaminidase liberated less than 1mole of N-acetylglucosamine and made at least 1mole further of mannose accessible to alpha-mannosidase action. 6. It is concluded that the carbohydrate moiety of ovalbumin is linked through a glycosyl group to asparagine. In a molecule with mannose/hexosamine ratio 5:4, there are two beta-N-acetylglucosamine residues linked together in a terminal position, followed by alpha-mannose. There is also present a side chain containing two alpha-mannose units.
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PMID:The enzymic degradation of ovalbumin and its glycopeptides. 535 18

We previously showed that under defined conditions beta-[3H]funaltrexamine (beta-[3H]FNA) covalently labeled mu-opioid receptors with high specificity in bovine striatal membranes. beta-[3H]FNA-labeled mu-opioid receptors migrated as a broad band with a molecular mass range of 68-97 kDa. It is controversial whether beta-FNA binds irreversibly to mu-opioid receptors in other species. In this study, we demonstrated that beta-[3H]FNA also labeled mu-opioid receptors with high specificity in brain membranes of the guinea pig, rat, and mouse. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and fluorography revealed that in each species beta-[3H]FNA specifically bound to a protein in which labeling was greatly reduced by naloxone. These labeled receptors had broad molecular mass ranges, and the molecular masses were different among these species, in the order of cow > guinea pig > rat > mouse. Membranes were subjected to solubilization with 2% Triton X-100 and wheat germ lectin (WGL) affinity chromatography. N-Acetylglucosamine eluted a peak of radioactivity. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and fluorography showed that in all four species the mu receptor was the only protein labeled with beta-[3H]FNA in the WGL eluate. The molecular masses of labeled mu-opioid receptors were 70-88 kDa (median, 77 kDa) for the cow, 66-80 kDa (median, 72 kDa) for the guinea pig, 60-75 kDa (median, 67 kDa) for the rat, and 60-72 kDa (median, 66 kDa) for the mouse. In addition, we investigated the nature of the carbohydrate moieties linked to the receptor protein and whether the species variation in the molecular mass was due to variable degrees of glycosylation. The bovine WGL eluate was treated with various glycosidases. Neuraminidase treatment decreased the receptor molecular mass by 6-7 kDa, whereas alpha-mannosidase had no effect. Removal of N-linked carbohydrates at asparagine residues by peptide-N4-[N-acetyl-beta-glucosaminyl]asparagine amidase (N-Glycanase) resulted in a much sharper specifically labelled protein band of 43 kDa. These results indicate that mu-opioid receptors are heavily glycosylated and the major carbohydrate moieties are of the complex type, N-linked to asparagine. After the WGL eluates for the four species were treated with N-Glycanase, the labeled receptors became much sharper bands with very similar molecular masses, i.e., 43 kDa for the cow and guinea pig, 39 kDa for the rat, and and 40 kDa for the mouse.(ABSTRACT TRUNCATED AT 400 WORDS)
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PMID:Beta-[3H]funaltrexamine-labeled mu-opioid receptors: species variations in molecular mass and glycosylation by complex-type, N-linked oligosaccharides. 823 25

Aspergillus fumigatus, the main etiological agent for various forms of human aspergillosis, gets access to the respiratory system of human host by inhalation of airborne conidia. These conidia possibly adhere to extracellular matrix (ECM) proteins. Among the ECM proteins involved in adherence, fibrinogen is thought to be crucial. Here, we studied whether A. fumigatus three-week culture filtrate (3wcf) proteins promote binding of A. fumigatus to ECM proteins and promote fungal growth. We observed that incubation of ECM with 3wcf proteins led to dose- and time-dependent increase in adherence of conidia to the ECM. In order to identify the catalogue of fibrinogen-binding A. fumigatus proteins, we carried out fibrinogen affinity blotting using two-dimensional gel electrophoresed 3wcf proteins. A total of 15 fibrinogen-binding protein spots corresponding to 7 unique proteins were identified in 3wcf using matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF-TOF). Among these, 4 proteins, namely, beta-glucosidase, alpha-mannosidase, pectate lyase A and oryzin precursor were predicted to have cell wall or extracellular localization, whereas amidase family protein and two hypothetical proteins did not display the signal sequence. This study reports seven novel fibrinogen-binding proteins of A. fumigatus, some of which could be further explored for targeting the adhesion phenomenon as antifungal strategy.
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PMID:Identification of fibrinogen-binding proteins of Aspergillus fumigatus using proteomic approach. 2187 Jan 22