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)

Formamidase from rat liver proved to be microheterogenous. After preparative isoelectric focusing in density gradient columns, two peaks of formamidase with identical substrate specificity were identified. By analytical focusing in thin layers of polyacrylamide or Sephadex G-75 SF, even five bands could be separated. Their isoelectric points were 4.75, 4.78, 4.82, 4.92 (main band) and 5.11, but their Michaelis constants did not differ significantly (54 to 62 mumol/l). An identical molecular weight of 34700 +/- 3200 for all bands was determined by disc electrophoresis. This value was confirmed by sedimentation analyses (so20,w = 3.00 S) and electrophoresis in the presence of sodium dodecyl-sulfate (Mr 34900 +/- 2300), which only gave a single band. The homogeneity was also confirmed by electrophoresis in the presence of 6M urea. Repeated disc electrophoresis of focusing under native conditions with single, isolated formamidases again resulted in different bands which were identified, not only by Coomassie Blue, but also by their hydrolytic cleavage of naphthyl acetate. Formamidase showed neither proteolytic nor asparagine-amidohydrolase activity and oligosaccharide conjugates were not detectable. Ampholytes, buffer ions, pH and peroxodisulfate did not affect the heterogeneity. "Initial burst" measurements with diethyl(4-nitrophenyl) phosphate yielded an equivalent weight of 36,300. Formylkynurenine reduced this inhibition very effectively. Thus, an extraordinary reactive serine residue appeared to be located in the catalytic site of formamidase. A participation of sulfhydrylgroups in the inactivating reaction of arsenite was excluded although two such groups were detected by 5,5'-dithiobis(2-nitrobenzoic acid). N-Bromosuccinimide reacted primarily with one of the nine tryptophan residues without loss of enzymatic activity, but a 18.6-fold excess of this reagent resulted in a complete loss of activity. The reaction rates of the most effective inhibitors and of the protective action of formylkynurenine were determined. Thus, formamidase must clearly be distinguished from typical serine esterases and proteases.
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PMID:[Formamidase--microheterogeneity, catalytic properties and inhibitors (author's transl)]. 8 81

To compare the value of spot urine and overnight 9-hour urine in the estimation of 24-hour urinary sodium excretion (UNaV), protein excretion (UpV) and kallikrein excretion (UKaV), we measured the concentration of sodium, protein, kallikrein and creatinine in spot urine, overnight 9-hour urine, and 24-hour urine samples obtained from 21 patients with various renal diseases. They ranged in age from 16 to 75 years with 10 males and 11 females. Urinary protein was measured by the Coomassie Blue dye-binding method. Urinary kallikrein activity was measured by assay of its amidase activity on synthetic substrate S-2266. The results showed that the 9-hour UpV and 9-hour urine P/Cr ratio was better correlated with the 24-hour UpV than the spot urine P/Cr ratio (at 9-11 AM), and the 9-hour UKaV and spot urine Ka/Cr ratio were better correlated with the 24-hour UKaV than the 9-hour Ka/Cr ratio. Only the 9-hour UNaV was correlated with the 24-hour UNaV. We conclude that overnight 9-hour urine, in view of its lower cost, equal effectiveness and convenience, is the best method to substitute for 24-hour urine collection in the evaluation of Na, P and Ka excretion in patients with renal diseases.
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PMID:Comparison between spot urine and overnight urine in the estimation of 24-hour excretion of urine protein, sodium and kallikrein. 168 68

The effects of zona pellucida glycoproteins, sulfated polymers and non-sulfated polymers on the activation kinetics of boar sperm proacrosin to beta-acrosin have been investigated. The aim has been to understand more about the behaviour and function of this protein after it has been released from the acrosome at the time of fertilization. Purified proacrosin was allowed to autoactivate at pH 8.0 in the presence of different concentrations of homologous zona glycoproteins, sulfated polymers (fucoidan, chondroitin sulfates A, B and C, dextran sulfate, polyvinylsulfate and heparin) and non-sulfated polymers (dextran, polyvinylphosphate and hyaluronic acid). Enzyme activity was measured against N-benzoyl-L-arginine p-nitroanalide substrate and changes in molecular mass of the protein monitored by SDS-PAGE. Results show that zona pellucida glycoproteins, fucoidan, dextran sulfate and polyvinylsulfate all potentiate the conversion of proacrosin to beta-acrosin but subsequently inhibit its amidase activity. Dextran, polyvinylphosphate, chondroitin sulfates A, B and C and glucose-6-sulfate, on the other hand, either have no effect on autoactivation and beta-acrosin activity, or enhance it slightly. SDS-PAGE analysis confirmed these observations and further indicated that binding of sulfated polymers to proacrosin inhibited staining by Coomassie Blue. These results are consistent with the hypothesis that binding of zona pellucida glycoproteins and sulfated compounds to proacrosin/acrosin is stereospecific and that contact activation onto soluble 'surfaces' causes conformational changes that are responsible for potentiation or inhibition of activation. The implications of these findings for sperm binding and penetration of the zona pellucida are discussed.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:Some effects of zona pellucida glycoproteins and sulfated polymers on the autoactivation of boar sperm proacrosin and activity of beta-acrosin. 818 87

We have developed an intermediate method toward the complete carbohydrate analysis of proteins, which should be universally applicable to all proteins and independent of sample matrix. Using only Coomassie Blue-stained proteins which have been electroblotted onto polyvinylidene fluoride membranes, we report a strategy for: (i) determining unequivocally whether a protein is glycosylated; (ii) obtaining a complete monosaccharide composition; (iii) oligosaccharide mapping which separates most forms according to size, charge and isomerity; and (iv) sequentially releasing and analyzing specific classes of oligosaccharides with endoglycosidases. The method was shown to be applicable to a variety of well characterized soluble glycoproteins and to the membrane-bound protein, the gastric H+, K(+)-ATPase. The monosaccharide composition of the H+,K(+)-ATPase revealed the absence of N-acetylneuraminic or N-glycolylneuraminic acids and a monosaccharide composition which indicated O-linked sugar chains. Oligomannosidic/hybrid and biantennary oligosaccharides were sequentially released and analyzed from one electroblotted band of recombinant tissue plasminogen activator using endo-beta-N-acetylglucosaminidase H and endo-beta-N-acetylglucosaminidase F2, respectively. Sialylated polylactosamine structures were identified and quantified by analyzing high performance liquid chromatography profiles of oligosaccharides first released by peptide-N4-(N-acetyl-beta-D-glucosaminyl)asparagine amidase and then treated with endo-beta-galactosidase, using a single, stained band of recombinant erythropoietin. This recombinant erythropoietin was found to contain eight times more tetrasialylated oligosaccharides than previously reported (Sasaki, H., Bothner, B., Dell, A., and Fukuda, M. (1987) J. Biol. Chem. 262, 12059-12076); 47% of released oligosaccharides were identified as polylactosamine structures.
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PMID:Monosaccharide and oligosaccharide analysis of proteins transferred to polyvinylidene fluoride membranes after sodium dodecyl sulfate-polyacrylamide gel electrophoresis. 844 88