Biomaterial Database

Cytosolic 5'-nucleotidase/nucleoside phosphotransferase: a single assay for a bifunctional enzyme. 1993

S Allegrini, and R Pesi, and M G Tozzi, and P L Ipata, and M Camici

Dipartimento di Fisiologia e Biochimica, Università di Pisa, Italy.

Cytosolic 5'-nucleotidase/nucleoside phosphotransferase has been purified from calf thymus. Since the same protein is able to catalyze both the hydrolysis and the interconversion of several nucleoside monophosphates, it is necessary to study the effect of different metabolites and assay conditions on both activities in order to elucidate their physiological roles. We describe herein a method which allowed us to follow both activities contemporaneously in the same assay mixture. The method takes advantage of the observation that deoxyGMP is both a good substrate for hydrolysis and a good phosphate donor for the phosphotransferase reaction, but its dephosphorylated product, deoxyguanosine, is not a phosphate acceptor. As a consequence, it is possible to follow both the deoxyguanosine production and the transfer of phosphate from deoxyGMP to the best phosphate acceptor, inosine, during the reaction, applying a method for the chromatographic separation on HPLC of both substrates (inosine and deoxyGMP) and both products (IMP and deoxyguanosine). The method was applied to the determination of the KM for inosine.

UI	MeSH Term	Description	Entries
D007291	Inosine Monophosphate	Inosine 5'-Monophosphate. A purine nucleotide which has hypoxanthine as the base and one phosphate group esterified to the sugar moiety.	IMP,Inosinic Acid,Ribosylhypoxanthine Monophosphate,Inosinic Acids,Sodium Inosinate,Acid, Inosinic,Acids, Inosinic,Inosinate, Sodium,Monophosphate, Inosine,Monophosphate, Ribosylhypoxanthine
D008722	Methods	A series of steps taken in order to conduct research.	Techniques,Methodological Studies,Methodological Study,Procedures,Studies, Methodological,Study, Methodological,Method,Procedure,Technique
D010770	Phosphotransferases	A rather large group of enzymes comprising not only those transferring phosphate but also diphosphate, nucleotidyl residues, and others. These have also been subdivided according to the acceptor group. (From Enzyme Nomenclature, 1992) EC 2.7.	Kinases,Phosphotransferase,Phosphotransferases, ATP,Transphosphorylase,Transphosphorylases,Kinase,ATP Phosphotransferases
D002417	Cattle	Domesticated bovine animals of the genus Bos, usually kept on a farm or ranch and used for the production of meat or dairy products or for heavy labor.	Beef Cow,Bos grunniens,Bos indicus,Bos indicus Cattle,Bos taurus,Cow,Cow, Domestic,Dairy Cow,Holstein Cow,Indicine Cattle,Taurine Cattle,Taurus Cattle,Yak,Zebu,Beef Cows,Bos indicus Cattles,Cattle, Bos indicus,Cattle, Indicine,Cattle, Taurine,Cattle, Taurus,Cattles, Bos indicus,Cattles, Indicine,Cattles, Taurine,Cattles, Taurus,Cow, Beef,Cow, Dairy,Cow, Holstein,Cows,Dairy Cows,Domestic Cow,Domestic Cows,Indicine Cattles,Taurine Cattles,Taurus Cattles,Yaks,Zebus
D002851	Chromatography, High Pressure Liquid	Liquid chromatographic techniques which feature high inlet pressures, high sensitivity, and high speed.	Chromatography, High Performance Liquid,Chromatography, High Speed Liquid,Chromatography, Liquid, High Pressure,HPLC,High Performance Liquid Chromatography,High-Performance Liquid Chromatography,UPLC,Ultra Performance Liquid Chromatography,Chromatography, High-Performance Liquid,High-Performance Liquid Chromatographies,Liquid Chromatography, High-Performance
D003600	Cytosol	Intracellular fluid from the cytoplasm after removal of ORGANELLES and other insoluble cytoplasmic components.	Cytosols
D006157	Guanosine Monophosphate	A guanine nucleotide containing one phosphate group esterified to the sugar moiety and found widely in nature.	5'-Guanylic Acid,Guanosine 5'-Monophosphate,5'-GMP,Guanylic Acid,5' Guanylic Acid,5'-Monophosphate, Guanosine,Acid, 5'-Guanylic,Acid, Guanylic,Guanosine 5' Monophosphate,Monophosphate, Guanosine
D006867	Hydrolases	Any member of the class of enzymes that catalyze the cleavage of the substrate and the addition of water to the resulting molecules, e.g., ESTERASES, glycosidases (GLYCOSIDE HYDROLASES), lipases, NUCLEOTIDASES, peptidases (PEPTIDE HYDROLASES), and phosphatases (PHOSPHORIC MONOESTER HYDROLASES). EC 3.	Hydrolase
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
D015720	5'-Nucleotidase	A glycoprotein enzyme present in various organs and in many cells. The enzyme catalyzes the hydrolysis of a 5'-ribonucleotide to a ribonucleoside and orthophosphate in the presence of water. It is cation-dependent and exists in a membrane-bound and soluble form. EC 3.1.3.5.	5'-AMP Nucleotidase,AMP Phosphatase,Adenylate Phosphatase,Antigens, CD73,CD73 Antigens,Cytidylate Phosphatase,Ecto-5'-Nucleotidase,IMP Nucleotidase,IMP Phosphatase,Inosinate Phosphatase,Pyrimidine 5'-Nucleotidase,Thymidine Phosphatase,Uridylate 5'-Nucleotidase,5'-Nucleotidase Phosphoribolase,Antigen, CD73,IMPase,5' AMP Nucleotidase,5' Nucleotidase,5' Nucleotidase Phosphoribolase,CD73 Antigen,Ecto 5' Nucleotidase,Pyrimidine 5' Nucleotidase,Uridylate 5' Nucleotidase