A poly(ADP-ribose) polymerase-1 activity assay based on the FRET between a cationic conjugated polymer and supercharged green fluorescent protein

Abstract

A label-free and convenient strategy for PARP-1 activity assay and inhibitors assessment has been developed based on the fluorescence resonance energy transfer (FRET) between a cationic conjugated polymer (CCP) and supercharged green fluorescent protein (scGFP).

Quartz Crystal Microbalance Detection of Poly(ADP-ribose) Polymerase-1 Based on Gold Nanorods Signal Amplification

“…2,5 The large, negatively charged PAR dramatically changes the charge properties of the acceptor proteins and initiates immediate cellular response to DNA damage. 6 Because PARP-1 activity level was related to tumor grade and the efficacy outcome of inhibitor therapy. 7−9 It is of great significance to develop some sensitive and accurate PARP-1 activity detection method for clinical diagnosis and drug screening in the treatment of various cancers.…”

“…Tang et al reported a convenient strategy for PARP-1 and its inhibitors assessment on the basis of the fluorescence resonance energy transfer between a cationic conjugated polymer and negatively supercharged green fluorescent protein. 6 Recently, our group also developed a simple and sensitive colorimetric biosensor based on the negatively charged PAR inducing the aggregation of cetyltrimethylammonium bromide (CTAB)-coated gold nanorods (GNRs). When PAR formed by PARP-1 was added, an obvious decrease of absorbance and vivid color change were observed.…”

“…As a ubiquitous and abundant nuclear enzyme, poly­(ADP-ribose) polymerase-1 (PARP-1) participates in many important cell processes including DNA repair and apoptosis. It plays a crucial role in tumorigenesis and other diseases. − Once activated by damaged DNA, quadruplexes, or specific dsDNA, PARP-1 cleaves nicotinamide adenine dinucleotide (NAD + ) and polymerizes the resulting ADP-ribose moiety (50–200 molecules) onto target proteins, ultimately forming a hyperbranched poly­(ADP-ribose) polymer (PAR). , The large, negatively charged PAR dramatically changes the charge properties of the acceptor proteins and initiates immediate cellular response to DNA damage . Because PARP-1 activity level was related to tumor grade and the efficacy outcome of inhibitor therapy. − It is of great significance to develop some sensitive and accurate PARP-1 activity detection method for clinical diagnosis and drug screening in the treatment of various cancers. − …”

“…Unfortunately, most of these assays needed sophisticated label procedures and the costs were expensive. Thus, some label-free and convenient strategy fluorescence techniques ,, and enzyme-free colorimetric assay , as well as electrochemical method , have been developed to overcome these shortcomings. Tang et al reported a convenient strategy for PARP-1 and its inhibitors assessment on the basis of the fluorescence resonance energy transfer between a cationic conjugated polymer and negatively supercharged green fluorescent protein .…”

Telomerase and poly(ADP-ribose) polymerase-1 activity sensing based on the high fluorescence selectivity and sensitivity of TOTO-1 towards G bases in single-stranded DNA and poly(ADP-ribose)

“…Most detection methods for PARP-1 are based on its catalyzed production of PAR that has a large number of negative charges, which can adsorb positively charged probes by electrostatic attraction 13–15. These methods do not have good selectivity because of the high background signals.…”