Premium
Carbon Nanotube‐Enhanced Polarization of Fluorescent Peptides: A Novel Amplification Strategy for Homogeneous Detection of Proteases
Author(s) -
Huang Yong,
Hu Kun,
Zhao Shulin,
Li Mei,
Chen ZhenFeng,
Lv Qiuju,
Liang Hong
Publication year - 2014
Publication title -
chemistry – an asian journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.18
H-Index - 106
eISSN - 1861-471X
pISSN - 1861-4728
DOI - 10.1002/asia.201301161
Subject(s) - fluorophore , proteases , protease , fluorescence anisotropy , peptide , carbon nanotube , fluorescence , chemistry , detection limit , biophysics , histidine , combinatorial chemistry , biochemistry , enzyme , nanotechnology , materials science , chromatography , biology , physics , quantum mechanics , membrane
Abstract A new fluorescence polarization (FP) amplification strategy based on the use of multiwalled carbon nanotubes (MWCNTs) as the FP enhancer was developed for the simple, sensitive, and universal monitoring of protease activity in homogeneous solution. A fluorophore‐labeled peptide that includes a protease‐cleavable element and ten histidine residues for binding MWCNTs is adsorbed on MWCNTs through strong π–π stacking and electrostatic interactions. When the fluorophore‐labeled peptide/MWCNT complexes are exposed to a protease target, specific peptide cleavage by the protease target occurs, thus releasing fragments carrying the fluorophore from the surface of MWCNTs, which in turn results in a significant decrease in the FP value. The detection limits of this assay for two proteases, thrombin and chymotrypsin (CTP), were estimated to be 0.5 p M and 0.3 p M , respectively. In addition, it is also demonstrated that this MWCNT‐enhanced FP assay is suitable for protease inhibitor screening.