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Detection of 3'→5' exonuclease activity using a metal-based luminescent switch-on probe
He H.-Z.1; Chan W.-I.2; Mak T.-Y.1; Liu L.-J.2; Wang M.1; Chan D.S.H.1; Ma D.-L.1; Leung C.-H.2
2013-12-15
Source PublicationMethods
ISSN10462023 10959130
Volume64Issue:3Pages:218-223
Abstract

A luminescent iridium(III) complex has been discovered to be selective for G-quadruplex DNA, and was employed in a label-free G-quadruplex-based detection assay for 3′ → 5′ exonuclease activity in aqueous solution. A proof-of-concept of this assay has been demonstrated by using prokaryotic exonuclease III (ExoIII) as a model enzyme. In this assay, a G-quadruplex-forming hairpin oligonucleotide (hairpin-G4 DNA, 5′-GAG3TG4AG3TG4A2GCAGA2G2ATA2CT2C4AC3TC4AC3TC-3′) initially exists in a duplex conformation, resulting in a low luminescence signal due to the weak interaction between the iridium(III) complex and duplex DNA. Upon digestion by ExoIII, the guanine-rich sequence is released and folds into a G-quadruplex, which greatly enhances the luminescence emission of the iridium(III) probe. This method was highly sensitive for 3′ → 5′ exonuclease over other DNA-modifying enzymes.

KeywordExonuclease G-quadruplex Iridium Luminescence Metal Complex
DOI10.1016/j.ymeth.2013.08.011
URLView the original
Indexed BySCIE
Language英语
WOS Research AreaBiochemistry & Molecular Biology
WOS SubjectBiochemical Research Methods ; Biochemistry & Molecular Biology
WOS IDWOS:000328309600006
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Cited Times [WOS]:16   [WOS Record]     [Related Records in WOS]
Document TypeJournal article
CollectionInstitute of Chinese Medical Sciences
Corresponding AuthorMa D.-L.; Leung C.-H.
Affiliation1.Department of Chemistry, Hong Kong Baptist University, Kowloon Tong, Hong Kong, China
2.State Key Laboratory of Quality Research in Chinese Medicine, Institute of Chinese Medical Sciences, University of Macau, Macao, China
Corresponding Author AffilicationInstitute of Chinese Medical Sciences
Recommended Citation
GB/T 7714
He H.-Z.,Chan W.-I.,Mak T.-Y.,et al. Detection of 3'→5' exonuclease activity using a metal-based luminescent switch-on probe[J]. Methods,2013,64(3):218-223.
APA He H.-Z..,Chan W.-I..,Mak T.-Y..,Liu L.-J..,Wang M..,...&Leung C.-H..(2013).Detection of 3'→5' exonuclease activity using a metal-based luminescent switch-on probe.Methods,64(3),218-223.
MLA He H.-Z.,et al."Detection of 3'→5' exonuclease activity using a metal-based luminescent switch-on probe".Methods 64.3(2013):218-223.
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