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A new highly sensitive and selective chemical assay for poly(ADP-ribose)

✍ Scribed by James L. Sims; Hector Juarez-Salinas; Myron K. Jacobson

Publisher
Elsevier Science
Year
1980
Tongue
English
Weight
909 KB
Volume
106
Category
Article
ISSN
0003-2697

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✦ Synopsis

A new method for the measurement of poly(ADP-ribose) is described. Poly(ADP-ribose) was separated from the bulk of cellular RNA and DNA by quantitative adsorption to dihydroxyboryl-Sepharose.

The polymer was digested with venom phosphodiesterase and bacterial alkaline phosphatase to yield the unique nucleoside 2' -+ 1" ribosyladenosine from internal residues. This nucleoside was converted by reaction with chloroacetaldehyde to a highly fluorescent I-N6-etheno derivative which was separated from interfering substances by reversed phase high-pressure liquid chromatography and picomole amounts were quantified by fluorescence. Control experiments are presented which demonstrate that the assay is highly specific for poly(ADP-ribose).

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