• 文献标题:   A Mix-and-Read Fluorescence Strategy for the Switch-On Probing of Kinase Activity Based on an Aptameric-Peptide/Graphene-Oxide Platform
  • 文献类型:   Article
  • 作  者:   LEI CY, XU XH, ZHOU J, LIU X, NIE Z, QING M, LI P, HUANG Y, YAO SZ
  • 作者关键词:   biosensor, fluorescence, graphene, kinase, peptide
  • 出版物名称:   CHEMISTRYAN ASIAN JOURNAL
  • ISSN:   1861-4728 EI 1861-471X
  • 通讯作者地址:   Hunan Univ
  • 被引频次:   4
  • DOI:   10.1002/asia.201402221
  • 出版年:   2014

▎ 摘  要

Protein kinase plays a vital role in regulating signal-transduction pathways and its simple and quick detection is highly desirable because traditional kinase assays typically rely on a time-consuming kinase-phosphorylation process (ca. 1 h). Herein, we report a new and rapid fluorescence-based sensing platform for probing the activity of protein kinase that is based on the super-quenching capacity of graphene oxide (GO) nanosheets and specific recognition of the aptameric peptide (FITC-IP20). On the GO/peptide platform, the fluorescence quenching of FITC-IP20 that is adsorbed onto GO can be restored by selective binding of active protein kinase to the aptameric peptide, thereby resulting in the fast switch-on detection of kinase activity (ca. 15 min). The feasibility of this method has been demonstrated by the sensitive measurement of the activity of cAMP-dependent protein kinase (PKA), with a detection limit of 0.053 mU mu L-1. This assay technique was also successfully applied to the detection of kinase activation in cell lysate.