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dc.contributor.authorWang, Chung-Liangen_US
dc.contributor.authorWei, Lan-Yien_US
dc.contributor.authorYuan, Chiun-Jyeen_US
dc.contributor.authorHwang, Kuo Chuen_US
dc.date.accessioned2014-12-08T15:21:28Z-
dc.date.available2014-12-08T15:21:28Z-
dc.date.issued2012-01-17en_US
dc.identifier.issn0003-2700en_US
dc.identifier.urihttp://dx.doi.org/10.1021/ac202369den_US
dc.identifier.urihttp://hdl.handle.net/11536/15253-
dc.description.abstractThis work presents a simple, low-cost and reusable label-free method for detecting protein tyrosine kinase activity using a tyrosinase-based amperometric biosensor (tyrosine kinase biosensor). This method is based on the observation that phosphorylation can block the tyrosinase-catalyzed oxidation of tyrosine or tyrosyl residue in peptides. Therefore, the activity of p60c-src protein tyrosine kinase (Src) on the developed tyrosine kinase biosensor could be quickly determined when its specific peptide substrate, p60c-src substrate I, was used. The tyrosine kinase biosensor was highly sensitive to the activity of Src with a linear dynamic range of 1.9-237.6 U/mL and the lowest detection limit of 0.23 U/mL. Interestingly, the tyrosine kinase activity can be measured using the developed tyrosine kinase biosensor repetitively without regeneration. The inhibitory effect of various kinase inhibitors on the Src activity could be determined on the tyrosine kinase biosensor. Src-specific inhibitors, PP2 and Src inhibitor I, effectively suppressed Src activity, whereas PD153035, an inhibitor of the epidermal growth factor receptor, was ineffective. Staurosporine, a universal kinase inhibitor, inhibited Src activity in an ATP concentration-dependent manner. These results suggests that the activities of tyrosine kinases and their behaviors toward various reagents can be effectively measured using the developed tyrosine kinase biosensor.en_US
dc.language.isoen_USen_US
dc.titleReusable Amperometric Biosensor for Measuring Protein Tyrosine Kinase Activityen_US
dc.typeArticleen_US
dc.identifier.doi10.1021/ac202369den_US
dc.identifier.journalANALYTICAL CHEMISTRYen_US
dc.citation.volume84en_US
dc.citation.issue2en_US
dc.citation.spage971en_US
dc.citation.epage977en_US
dc.contributor.department生物科技學系zh_TW
dc.contributor.department分子醫學與生物工程研究所zh_TW
dc.contributor.departmentDepartment of Biological Science and Technologyen_US
dc.contributor.departmentInstitute of Molecular Medicine and Bioengineeringen_US
dc.identifier.wosnumberWOS:000299134400036-
dc.citation.woscount13-
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