Development of a fluorescence peptide chip for the detection of caspase activity.
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.
Han
Aishan
A
Department of Applied Chemistry, Faculty of Engineering, Kyushu University, 6-10-1 Hakozaki, Higashi-Ku, Fukuoka 812-8581, Japan.
Sonoda
Tatsuhiko
T
Kang
Jeong-Hun
JH
Murata
Masaharu
M
NIiidome
Takuro
T
Katayam
Yoshiki
Y
D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D016466:CHO Cells; D053148:Caspase 3; D020169:Caspases / Q000737:chemistry* / Q000378:metabolism*; D002478:Cells, Cultured; D006224:Cricetinae; D004789:Enzyme Activation; D005456:Fluorescent Dyes / Q000737:chemistry*; D008856:Microscopy, Fluorescence; D015394:Molecular Structure; D010455:Peptides / Q000737:chemistry*; D040081:Protein Array Analysis / Q000379:methods*; D012680:Sensitivity and Specificity; D019311:Staurosporine / Q000494:pharmacology; D013499:Surface Properties; D013997:Time Factors
D005456:Fluorescent Dyes; D010455:Peptides; D053148:Caspase 3; D020169:Caspases; D019311:Staurosporine