標題: Reusable Amperometric Biosensor for Measuring Protein Tyrosine Kinase Activity
作者: Wang, Chung-Liang
Wei, Lan-Yi
Yuan, Chiun-Jye
Hwang, Kuo Chu
生物科技學系
分子醫學與生物工程研究所
Department of Biological Science and Technology
Institute of Molecular Medicine and Bioengineering
公開日期: 17-一月-2012
摘要: This 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.
URI: http://dx.doi.org/10.1021/ac202369d
http://hdl.handle.net/11536/15253
ISSN: 0003-2700
DOI: 10.1021/ac202369d
期刊: ANALYTICAL CHEMISTRY
Volume: 84
Issue: 2
起始頁: 971
結束頁: 977
顯示於類別:期刊論文


文件中的檔案:

  1. 000299134400036.pdf