Label-free detection of protein-protein interactions using a calmodulin-modified nanowire transistor.

نویسندگان

  • Tsung-Wu Lin
  • Po-Jen Hsieh
  • Chih-Lung Lin
  • Yi-Ya Fang
  • Jia-Xun Yang
  • Chia-Chang Tsai
  • Pei-Ling Chiang
  • Chien-Yuan Pan
  • Yit-Tsong Chen
چکیده

In this study, we describe a highly sensitive and reusable silicon nanowire field-effect transistor for the detection of protein-protein interactions. This reusable device was made possible by the reversible association of glutathione S-transferase-tagged calmodulin with a glutathione modified transistor. The calmodulin-modified transistor exhibited selective electrical responses to Ca2+ (> or = 1 microM) and purified cardiac troponin I (approximately 7 nM); the change in conductivity displayed a linear dependence on the concentration of troponin I in a range from 10 nM to 1 microM. These results are consistent with the previously reported concentration range in which the dissociation constant for the troponin I-calmodulin complex was determined. The minimum concentration of Ca2+ required to activate calmodulin was determined to be 1 microM. We have also successfully demonstrated that the N-type Ca2+ channels, expressed by cultured 293T cells, can be recognized specifically by the calmodulin-modified nanowire transistor. This sensitive nanowire transistor can serve as a high-throughput biosensor and can also substitute for immunoprecipitation methods used in the identification of interacting proteins.

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عنوان ژورنال:
  • Proceedings of the National Academy of Sciences of the United States of America

دوره 107 3  شماره 

صفحات  -

تاریخ انتشار 2010