Document Detail


Modified phosphoribosylpyrophosphate (PRPP) radioenzymatic assay: increased sensitivity, technical simplification and new applications.
MedLine Citation:
PMID:  2439241     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Phosphoribosylpyrophosphate in amounts as low as 25 pmol could be reliably and economically measured with a CO2-releasing radioenzymatic assay when appropriate technical modifications were introduced. The concentration of commercially available phosphoribosylpyrophosphate used for reference standards was ascertained by a method based on the utilization of phosphoribosylpyrophosphate by hypoxanthine catalyzed by hypoxanthine phosphoribosyltransferase from red blood cell lysates. The addition of inorganic phosphate increased intracellular phosphoribosylpyrophosphate levels in HL-60 cell lysates and can be used to amplify low levels of phosphoribosylpyrophosphate. This phosphoribosylpyrophosphate assay amplified by inorganic phosphate has been developed to assay perturbations in the purine biosynthetic nucleotide pathway in response to various chemotherapeutic agents, such as anti-folates, or as a result of folate deficiency.
Authors:
J Ghitis; S Waxman
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Clinica chimica acta; international journal of clinical chemistry     Volume:  164     ISSN:  0009-8981     ISO Abbreviation:  Clin. Chim. Acta     Publication Date:  1987 May 
Date Detail:
Created Date:  1987-08-17     Completed Date:  1987-08-17     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  1302422     Medline TA:  Clin Chim Acta     Country:  NETHERLANDS    
Other Details:
Languages:  eng     Pagination:  267-73     Citation Subset:  IM    
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MeSH Terms
Descriptor/Qualifier:
Cell Line
Erythrocytes / analysis
Humans
Hypoxanthine
Hypoxanthine Phosphoribosyltransferase / metabolism
Hypoxanthines
Pentosephosphates / analysis*
Phosphoribosyl Pyrophosphate / analysis*,  blood
Reference Standards
Chemical
Reg. No./Substance:
0/Hypoxanthines; 0/Pentosephosphates; 68-94-0/Hypoxanthine; 7540-64-9/Phosphoribosyl Pyrophosphate; EC 2.4.2.8/Hypoxanthine Phosphoribosyltransferase

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