PMID- 31774658
OWN - NLM
STAT- MEDLINE
DCOM- 20201027
LR  - 20201027
IS  - 1520-5126 (Electronic)
IS  - 0002-7863 (Print)
IS  - 0002-7863 (Linking)
VI  - 141
IP  - 49
DP  - 2019 Dec 11
TI  - Ultrafast Oxime Formation Enables Efficient Fluorescence Light-up Measurement of 
      DNA Base Excision.
PG  - 19379-19388
LID - 10.1021/jacs.9b09812 [doi]
AB  - DNA glycosylases constitute a biologically and biomedically important group of 
      DNA repair enzymes responsible for initiating base excision repair (BER). 
      Measuring their activities can be useful for studying the mechanisms DNA damage 
      and repair and for practical applications in cancer diagnosis and drug screening. 
      Previous fluorescence methods for assaying DNA glycosylases are often complex 
      and/or limited in scope to a single enzyme type. Here we report a universal base 
      excision reporter (UBER) fluorescence probe design that implements an 
      unprecedentedly rapid oxime reaction (>150 M(-1) s(-1)) with high specificity for 
      the abasic (AP) site of DNA. The molecular rotor design achieves a robust 
      >250-500-fold increase in fluorescence upon reaction with AP sites in DNA. By 
      using the fluorescence reporter in concert with specific DNA lesion-containing 
      substrates, the UBER probe can be used in a coupled assay in principle with any 
      DNA glycosylase. We demonstrate the utility of the UBER probe by assaying five 
      different glycosylases in real time as well as profiling glycosylase activity in 
      cell lysates. We anticipate that the UBER probe will be of considerable utility 
      to researchers studying DNA repair biology owing to its high level of 
      generalizability, ease of use, and compatibility with biologically derived 
      samples.
FAU - Wilson, David L
AU  - Wilson DL
AUID- ORCID: 0000-0002-1922-427X
AD  - Department of Chemistry, Stanford Cancer Institute and ChEM-H Institute , 
      Stanford University , Stanford , California 94305 , United States.
FAU - Kool, Eric T
AU  - Kool ET
AUID- ORCID: 0000-0002-7310-2935
AD  - Department of Chemistry, Stanford Cancer Institute and ChEM-H Institute , 
      Stanford University , Stanford , California 94305 , United States.
LA  - eng
GR  - R01 CA217809/CA/NCI NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, Non-U.S. Gov't
DEP - 20191127
PL  - United States
TA  - J Am Chem Soc
JT  - Journal of the American Chemical Society
JID - 7503056
RN  - 0 (Fluorescent Dyes)
RN  - 0 (Oximes)
RN  - 9007-49-2 (DNA)
RN  - EC 3.2.2.- (DNA Glycosylases)
SB  - IM
MH  - DNA/*chemistry
MH  - DNA Damage
MH  - DNA Glycosylases/*chemistry
MH  - *DNA Repair
MH  - Fluorescent Dyes/*chemistry
MH  - Fluorometry
MH  - Molecular Structure
MH  - Oximes/*chemistry
PMC - PMC7061905
MID - NIHMS1558496
COIS- The authors declare no competing financial interest.
EDAT- 2019/11/28 06:00
MHDA- 2020/10/28 06:00
PMCR- 2020/03/09
CRDT- 2019/11/28 06:00
PHST- 2019/11/28 06:00 [pubmed]
PHST- 2020/10/28 06:00 [medline]
PHST- 2019/11/28 06:00 [entrez]
PHST- 2020/03/09 00:00 [pmc-release]
AID - 10.1021/jacs.9b09812 [doi]
PST - ppublish
SO  - J Am Chem Soc. 2019 Dec 11;141(49):19379-19388. doi: 10.1021/jacs.9b09812. Epub 
      2019 Nov 27.