Development of a Fluorescence Peptide Chip for the Detection of Caspase Activity

Title: Development of a Fluorescence Peptide Chip for the Detection of Caspase Activity

Volume: 9 Issue: 1

Author(s): Aishan Han, Tatsuhiko Sonoda, Jeong-Hun Kang, Masaharu Murata, Takuro Niiidome and Yoshiki Katayama

Affiliation:

Keywords: Caspase-3, micro array, apoptosis, protease, FRET, cell lysate

Abstract: Proteases play a key role in cell functions, and it is very important to monitor their activities for drug screening and diagnosis of diseases. In the present study, a new class of fluorescence probe, into which a fluorophore and a quencher have been introduced, was developed and applied to the on-chip detection of caspase-3 activity. This probe is non fluorescent in the absence of caspase-3. However, when it is treated with active caspase-3, the fluorescence intensity increases dependent on the caspase-3 activity due to the cleavage of the quencher-containing moiety on a glass slide. This caspase-dependent increase in the fluorescence intensity was also detected when the glass slide immobilizing the probe peptide was treated with cell lysate stimulated by staurosporine (STP), which is an apoptosis-inducing agent. On the other hand, such an increase was not detected in the case of control cell lysate without STP-stimulation. The developed system is a rapid and sensitive method and is useful for the direct measurement of protease activity on a glass array.

Cite this article as:

Han Aishan, Sonoda Tatsuhiko, Kang Jeong-Hun, Murata Masaharu, Niiidome Takuro and Katayama Yoshiki, Development of a Fluorescence Peptide Chip for the Detection of Caspase Activity, Combinatorial Chemistry & High Throughput Screening 2006; 9(1) . https://dx.doi.org/10.2174/138620706775213831