A label-free real time fluorometric assay for protease and inhibitor screening with a released heme

A label free continuous assay for protease activity and inhibitor screening has been developed. A protease (trypsin) could digest hemoglobin. Free heme molecules were released. Strong π-π stacking and hydrophobic interactions with the perylene probe resulted in efficient quenching of the probe'...

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Bibliographic Details
Published in:Chemical communications (Cambridge, England) England), 2012-10, Vol.48 (81), p.10123-10125
Main Authors: Li, Wenying, Chen, Jian, Jiao, Huping, Zhang, Qingfeng, Zhou, Huipeng, Yu, Cong
Format: Article
Language:English
Subjects:
Animals
Cattle
Enzyme Assays - methods
Fluorescent Dyes - analysis
Fluorescent Dyes - metabolism
Heme - metabolism
Humans
Perylene - analysis
Perylene - metabolism
Sensitivity and Specificity
Spectrometry, Fluorescence - methods
Trypsin - metabolism
Trypsin Inhibitors - pharmacology
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Summary:A label free continuous assay for protease activity and inhibitor screening has been developed. A protease (trypsin) could digest hemoglobin. Free heme molecules were released. Strong π-π stacking and hydrophobic interactions with the perylene probe resulted in efficient quenching of the probe's monomer fluorescence.
ISSN:1359-7345
1364-548X
DOI:10.1039/c2cc35374a