In:
Proceedings of the National Academy of Sciences, Proceedings of the National Academy of Sciences, Vol. 97, No. 25 ( 2000-12-05), p. 13537-13542
Abstract:
Although extensive effort has been applied toward
understanding the mechanism by which enediynes cleave DNA, a continuous assay for this phenomenon is still lacking. In fact, with the exception
of assays for DNase, continuous assays for most DNA cleavage events are unavailable. This article describes the application of “molecular
break lights” (a single-stranded oligonucleotide that adopts a stem-and-loop structure and carries a 5′-fluorescent moiety, a
3′-nonfluorescent quenching moiety, and an appropriate cleavage site within the stem) to develop the first continuous assay for cleavage of
DNA by enediynes. Furthermore, the generality of this approach is demonstrated by using the described assay to directly compare the DNA
cleavage by naturally occurring enediynes [calicheamicin and esperamicin), non-enediyne small molecule agents (bleomycin,
methidiumpropyl–EDTA–Fe(II), and EDTA–Fe(II] ), as well as the
restriction endonuclease Bam HI. Given the simplicity,
speed, and sensitivity of this approach, the described methodology could easily be extended to a high throughput format and become a new
method of choice in modern drug discovery to screen for novel protein-based or small molecule-derived DNA cleavage agents.
Type of Medium:
Online Resource
ISSN:
0027-8424
,
1091-6490
DOI:
10.1073/pnas.240460997
Language:
English
Publisher:
Proceedings of the National Academy of Sciences
Publication Date:
2000
detail.hit.zdb_id:
209104-5
detail.hit.zdb_id:
1461794-8
SSG:
11
SSG:
12
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