Journal
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
Volume 107, Issue 3, Pages 1047-1052Publisher
NATL ACAD SCIENCES
DOI: 10.1073/pnas.0910243107
Keywords
calcium ion; glutathione S-transfrease; N-type calcium channel; silicon nanowire field-effect transistor
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Funding
- National Science Council of Taiwan [NSC 97-2627-M-002-019, NSC 97-2627-M-002-020]
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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+(>= 1 mu M) and purified cardiac troponin I (similar to 7 nM); the change in conductivity displayed a linear dependence on the concentration of troponin I in a range from 10 nM to 1 mu M. 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 mu M. 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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