ABSTRACT
The effect of ozone stress on polyamine metabolism and membrane lipid peroxidation in lentil seedlings through the amine oxidase and lipoxygenase activity and expression has been investigated. Ozone is shown to control the expression of these enzymes at the transcriptional level, down-regulating the amine oxidase gene and up-regulating the lipoxygenase gene. The decrease of amine oxidase activity correlated with the increase of putrescine concentration in the ozone-treated plantlets, whereas the increase of lipoxygenase activity was paralleled by enhanced membrane lipid peroxidation. Finally, polyamines are shown to inhibit lipoxygenase activity in lentils.
Subject(s)
Amine Oxidase (Copper-Containing) , Fabaceae/enzymology , Gene Expression Regulation, Plant/drug effects , Lipoxygenase/genetics , Oxidative Stress , Oxidoreductases Acting on CH-NH Group Donors/genetics , Ozone/pharmacology , Plants, Medicinal , Fabaceae/drug effects , Fabaceae/genetics , Lipid Peroxidation/drug effects , Lipid Peroxides/metabolism , Lipoxygenase/biosynthesis , Lipoxygenase Inhibitors/pharmacology , Membrane Lipids/metabolism , Oxidoreductases Acting on CH-NH Group Donors/biosynthesis , Polyamines/metabolism , Polyamines/pharmacology , Putrescine/metabolism , Putrescine/pharmacology , RNA, Messenger/genetics , RNA, Messenger/metabolism , Seeds/drug effects , Seeds/enzymology , Seeds/genetics , Spermidine/metabolism , Spermidine/pharmacology , Spermine/metabolism , Spermine/pharmacologyABSTRACT
Human Tamm-Horsfall glycoprotein has been purified from the urine of one male. The Asn-linked carbohydrate chains were enzymically released by peptide-N4-(N-acetyl-beta-glucosaminyl)asparagine amidase F, and separated from the remaining protein by gel-permeation chromatography on Bio-Gel P-100. Fractionation of the intact (sulfated) sialylated carbohydrate chains was achieved by a combination of three liquid-chromatographic techniques, namely, anion-exchange FPLC on Q-Sepharose, amine-adsorption HPLC on Lichrospher-NH2, and high-pH anion-exchange chromatography on CarboPac PA1. In total, more than 150 carbohydrate-containing fractions were obtained, some of which still contained mixtures of oligosaccharides. The primary structure of 30 N-glycans, including 10 novel oligosaccharides, were determined by one- and two-dimensional 1H-NMR spectroscopy at 500 MHz or 600 MHz. The types of compounds identified range from non-fucosylated, monosialylated, diantennary to fucosylated, tetrasialylated, tetraantennary carbohydrate chains, possessing the following terminal structural elements: [formula: see text]
Subject(s)
Acetylgalactosamine/chemistry , Asparagine/chemistry , Carbohydrates/chemistry , Mucoproteins/chemistry , Sulfates/chemistry , Carbohydrate Sequence , Chromatography, Ion Exchange , Chromatography, Liquid , Humans , Magnetic Resonance Spectroscopy , Male , Molecular Sequence Data , UromodulinABSTRACT
New born meconium contains at least a hundred oligosaccharides. In this study the isolation and characterization of the major constituents is described. The structure elucidation of 15 neutral and acidic oligosaccharides was carried out by methylation analysis, mass spectrometry and 360-MHz 1H-NMR spectroscopy. The results show that the oligosaccharides accumulating in human meconium are probably products of the catabolism of the O- and N-linked carbohydrate chains of glycoproteins. It is proposed that endo-N-acetyl-alpha-D-galactosaminidase, endo-beta-D-galactosidase and endo-N-acetyl-beta-D-glucosaminidase are involved in the production of these compounds.
Subject(s)
Meconium/analysis , Oligosaccharides/isolation & purification , Carbohydrate Conformation , Carbohydrate Sequence , Carbohydrates/analysis , Humans , Infant, Newborn , Magnetic Resonance Spectroscopy , Mass SpectrometryABSTRACT
The isolation, identification, and quantitative determination of an unusual urinary dipeptide, Nepsilon-(beta-aspartyl)lysine, is described, as well as its synthesis. This compound was observed in children in various disease states, but without any correlation with a particular symptom. Its origin is discussed.
Subject(s)
Dipeptides/urine , Aspartic Acid/urine , Child , Child, Preschool , Chromatography, Gas , Chromatography, Thin Layer , Female , Humans , Infant , Lysine/urine , Male , Mass SpectrometryABSTRACT
In addition to the already knownonosaccharides fucose, xylose, mannose, galactose, glucose, N-acetylgalactosamine and N-acetylglucosamine, the carbohydrate part of the haemocyanin from Helix pomatia (Roman snail) contains 3-O-methylgalactose, and that from Lymnaea stagnalis (a freshwater snail) 3-O-methylgalactose and 3-O-methylmannose. The 3-O-methyl sugars were identified by g.l.c.-mas spectrometry of the corresponding trimethylsilyl methyl glycosides and the alditol acetates, and by co-chromatography with the synthetic reference substances.
