Configuration of a scintillation proximity assay for the activity assessment of recombinant human adenine phosphoribosyltransferase.

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Barrett C, Alley J, Pulido JC, Spurling H, Li P, Parsons T, Mallender WD, Bembenek ME

Configuration of a scintillation proximity assay for the activity assessment of recombinant human adenine phosphoribosyltransferase.

Assay Drug Dev Technol. 2006 Dec;4(6):661-9.

PubMed ID

17199504 [ View in PubMed

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Abstract

Adenine phosphoribosyltransferase plays a role in purine salvage by catalyzing the direct conversion of adenine to adenosine monophosphate. The involvement of the purine salvage pathway in tumor proliferation and angiogenesis makes adenine phosphoribosyltransferase a potential target for oncology drug discovery. We have expressed and characterized recombinant, N-terminally His-tagged human adenine phosphoribosyltransferase. Two assay formats were assessed for use in a high throughput screen: a spectrophotometric-based enzyme-coupled assay system and a radiometric ionic capture scintillation proximity bead assay format. Ultimately, the scintillation proximity assay format was chosen because of automated screening compatibility limitations of the coupled assay. We describe here the biochemical characterization of adenine phosphoribosyltransferase and the development of a robust, homogeneous, 384-well assay suitable for high throughput screening.

DrugBank Data that Cites this Article

Drug Targets

Drug	Target	Kind	Organism	Pharmacological Action	Actions
Adenine	Adenine phosphoribosyltransferase	Protein	Humans	Unknown	Not Available	Details

Drug Enzymes

Drug	Enzyme	Kind	Organism	Pharmacological Action	Actions
Adenosine phosphate	Adenine phosphoribosyltransferase	Protein	Humans	Unknown	Substrate	Details