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

Article Details

Citation

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
]
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
DrugTargetKindOrganismPharmacological ActionActions
AdenineAdenine phosphoribosyltransferaseProteinHumans
Unknown
Not AvailableDetails
Drug Enzymes
DrugEnzymeKindOrganismPharmacological ActionActions
Adenosine phosphateAdenine phosphoribosyltransferaseProteinHumans
Unknown
Substrate
Details