A simplified method for estimating 5-phosphoribosyl 1-pyrophosphate in mouse liver and spleen. 1984

A H Chalmers

A simplified method for the estimation of 5-phosphoribosyl 1-pyrophosphate (PRPP) in mouse liver and spleen is described. The method uses the enzymic conversion of [8-14C] hypoxanthine to [8-14C] inosine 5'-monophosphate in the presence of PRPP and has the advantage over previously published methods in that the enzyme used in this assay, hypoxanthine-guanine phosphoribosyltransferase (HG-PRTase), is present in the tissue being analyzed and does not require preparation. In addition, each assay uses as its internal standard PRPP, thus overcoming the problem of the chemical instability inherent in this molecule. The two-minute heat treatment used in the method was found to destroy most interfering enzymes and resulted in average recovery of PRPP of 56% and 85% for the liver and spleen, respectively. The counts obtained in the assay were about ten-fold above background (80 c.p.m) and allowed accurate measurement of the phosphoribose sugar. The levels of PRPP (+/- standard deviation) found for liver and spleen in twelve (12) separate Balb/C male mice were 6.3 +/- 1.8 and 10.8 +/- 5.0 nmoles/g wet weight tissue, respectively, and agree with the tissue concentrations already reported for this metabolite. A further advantage of this assay is that it gives an index of the HG-PRTase activity in the tissues.

UI MeSH Term Description Entries
D007041 Hypoxanthine Phosphoribosyltransferase An enzyme that catalyzes the conversion of 5-phosphoribosyl-1-pyrophosphate and hypoxanthine, guanine, or MERCAPTOPURINE to the corresponding 5'-mononucleotides and pyrophosphate. The enzyme is important in purine biosynthesis as well as central nervous system functions. Complete lack of enzyme activity is associated with the LESCH-NYHAN SYNDROME, while partial deficiency results in overproduction of uric acid. EC 2.4.2.8. Guanine Phosphoribosyltransferase,HPRT,Hypoxanthine-Guanine Phosphoribosyltransferase,IMP Pyrophosphorylase,HGPRT,HPRTase,Hypoxanthine Guanine Phosphoribosyltransferase,Phosphoribosyltransferase, Guanine,Phosphoribosyltransferase, Hypoxanthine,Phosphoribosyltransferase, Hypoxanthine-Guanine,Pyrophosphorylase, IMP
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
D008297 Male Males
D008807 Mice, Inbred BALB C An inbred strain of mouse that is widely used in IMMUNOLOGY studies and cancer research. BALB C Mice, Inbred,BALB C Mouse, Inbred,Inbred BALB C Mice,Inbred BALB C Mouse,Mice, BALB C,Mouse, BALB C,Mouse, Inbred BALB C,BALB C Mice,BALB C Mouse
D010428 Pentosephosphates
D010754 Phosphoribosyl Pyrophosphate The key substance in the biosynthesis of histidine, tryptophan, and purine and pyrimidine nucleotides. Pyrophosphate, Phosphoribosyl
D006358 Hot Temperature Presence of warmth or heat or a temperature notably higher than an accustomed norm. Heat,Hot Temperatures,Temperature, Hot,Temperatures, Hot
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
D013154 Spleen An encapsulated lymphatic organ through which venous blood filters.
D051379 Mice The common name for the genus Mus. Mice, House,Mus,Mus musculus,Mice, Laboratory,Mouse,Mouse, House,Mouse, Laboratory,Mouse, Swiss,Mus domesticus,Mus musculus domesticus,Swiss Mice,House Mice,House Mouse,Laboratory Mice,Laboratory Mouse,Mice, Swiss,Swiss Mouse,domesticus, Mus musculus

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