Enzymatic determination of free glucuronic acid with glucuronolactone reductase. II. Procedure for the enzymatic determination of glucuronic acid and its application to degradation studies of glycosaminoglycans. 1984

S Hayashi, and M Watanabe, and A Kimura

An enzymatic method was devised for determination of free glucuronic acid by the use of glucuronolactone reductase from rat kidney. Free glucuronic acid in the range of 4 to 200 micrograms was determined quantitatively by this method even in the presence of neutral sugars and oligosaccharides of glycosaminoglycans. The interference due to 20 to 100 micrograms of aldohexose or aldopentose was less than 0.5% at the same concentration of free glucuronic acid. The activity of the enzyme towards substituted glucuronic acid was only 1% of that towards free glucuronic acid. The enzyme did not act on N-acetylhexosamines. The method was applied to a study of glycosaminoglycan degradation by lysosomal enzyme. When hyaluronic acid and chondroitin 4-sulfate were incubated for 24 h with a glycoprotein fraction obtained from rat liver lysosomes by concanavalin A agarose chromatography, free glucuronic acid was liberated (showing a sigmoid curve as a function of incubation time) and reached 21 and 10% of total glucuronic acid, respectively. Tetrasaccharide from chondroitin 4-sulfate was degraded to equimolar amounts of free glucuronic acid and sulfated trisaccharide.

UI MeSH Term Description Entries
D008099 Liver A large lobed glandular organ in the abdomen of vertebrates that is responsible for detoxification, metabolism, synthesis and storage of various substances. Livers
D008247 Lysosomes A class of morphologically heterogeneous cytoplasmic particles in animal and plant tissues characterized by their content of hydrolytic enzymes and the structure-linked latency of these enzymes. The intracellular functions of lysosomes depend on their lytic potential. The single unit membrane of the lysosome acts as a barrier between the enzymes enclosed in the lysosome and the external substrate. The activity of the enzymes contained in lysosomes is limited or nil unless the vesicle in which they are enclosed is ruptured or undergoes MEMBRANE FUSION. (From Rieger et al., Glossary of Genetics: Classical and Molecular, 5th ed). Autolysosome,Autolysosomes,Lysosome
D008297 Male Males
D008832 Microchemistry The development and use of techniques and equipment to study or perform chemical reactions, with small quantities of materials, frequently less than a milligram or a milliliter.
D002237 Carbohydrate Dehydrogenases Reversibly catalyze the oxidation of a hydroxyl group of carbohydrates to form a keto sugar, aldehyde or lactone. Any acceptor except molecular oxygen is permitted. Includes EC 1.1.1.; EC 1.1.2.; and 1.1.99. Carbohydrate Oxidoreductases,Dehydrogenases, Carbohydrate,Oxidoreductases, Carbohydrate
D005965 Glucuronates Derivatives of GLUCURONIC ACID. Included under this heading are a broad variety of acid forms, salts, esters, and amides that include the 6-carboxy glucose structure. Glucosiduronates,Glucuronic Acids,Acids, Glucuronic
D006025 Glycosaminoglycans Heteropolysaccharides which contain an N-acetylated hexosamine in a characteristic repeating disaccharide unit. The repeating structure of each disaccharide involves alternate 1,4- and 1,3-linkages consisting of either N-acetylglucosamine (see ACETYLGLUCOSAMINE) or N-acetylgalactosamine (see ACETYLGALACTOSAMINE). Glycosaminoglycan,Mucopolysaccharides
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia
D051381 Rats The common name for the genus Rattus. Rattus,Rats, Laboratory,Rats, Norway,Rattus norvegicus,Laboratory Rat,Laboratory Rats,Norway Rat,Norway Rats,Rat,Rat, Laboratory,Rat, Norway,norvegicus, Rattus
D020723 Glucuronic Acid A sugar acid formed by the oxidation of the C-6 carbon of GLUCOSE. In addition to being a key intermediate metabolite of the uronic acid pathway, glucuronic acid also plays a role in the detoxification of certain drugs and toxins by conjugating with them to form GLUCURONIDES. Glucuronate,Glucuronic Acid, 6-(14)C-labeled, (D)-isomer,Glucuronic Acid, Monopotassium Salt,Glucuronic Acid, Monosodium Salt,Monopotassium Glucuronate,Monosodium Glucuronate,Glucuronate, Monopotassium,Glucuronate, Monosodium

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