Surface plasmon resonance imaging measurements of caspase reactions on peptide microarrays.
Enzymatic activity monitoring of caspases, which are a class of cysteine protease, was performed by using peptide arrays based on surface plasmon resonance (SPR) imaging. The strategy of the detection is straightforward, using streptavidin to amplify the SPR signals of the surface-immobilized substrate peptides labeled with biotin at the C termini. Thus, the cleavage of the substrate peptides by caspases was detected as a signal decrease. Using this method, we succeeded in monitoring the activities of purified caspases and caspases in cell lysates. The SPR imaging-based peptide array would be applicable to cell-based drug screening and biochemical studies to reveal signal transduction processes.
Inoue
Yusuke
Y
Graduate School of Systems Life Sciences, Kyushu University, Fukuoka 819-0395, Japan.
Mori
Takeshi
T
Yamanouchi
Go
G
Han
Xiaoming
X
Sonoda
Tatsuhiko
T
Niidome
Takuro
T
Katayama
Yoshiki
Y
D000595:Amino Acid Sequence; D000818:Animals; D001710:Biotin / Q000378:metabolism; D016466:CHO Cells; D020169:Caspases / Q000378:metabolism*; D006224:Cricetinae; D003412:Cricetulus; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry; D040081:Protein Array Analysis / Q000379:methods*; D019311:Staurosporine / Q000494:pharmacology; D019809:Streptavidin / Q000378:metabolism; D020349:Surface Plasmon Resonance / Q000379:methods*
D010455:Peptides; D001710:Biotin; D019809:Streptavidin; D020169:Caspases; D019311:Staurosporine