Journal
SENSORS AND ACTUATORS B-CHEMICAL
Volume 232, Issue -, Pages 557-563Publisher
ELSEVIER SCIENCE SA
DOI: 10.1016/j.snb.2016.03.144
Keywords
Electrochemical sensors; Protease; ATCUN motif; Copper; beta-secretase
Funding
- National Natural Science Foundation of China [21205003, 21305004]
- Joint Fund for Fostering Talents of National Natural Science Foundation of China
- Henan Province [U1304205]
- Program for Science and Technology Innovation Talents at the University of Henan Province [15HASTIT001]
- Program for Innovative Research Team of Science and Technology in the University of Henan Province [16IRT-STHN003]
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Amino terminal Cu(II)- and Ni(II)-binding (ATCUN) motif is included in a protein or peptide with a structural characterization comprising of a free NH2-terminus, a histidine residue in the third position and two intervening peptide nitrogens. We reported a signal-on electrochemical strategy for detection of protease based on the formation of ATCUN-Cu(II) on electrode surface. In the method, an amino residue in the native peptide substrate was replaced with a histidine. Cleavage of the sequence-specific peptide pre-immobilized on an electrode by the protease led to the presence of ATCUN motif and the formation of ATCUN-Cu(II) in the presence of Cu(II). Consequently, a voltammetric signal was observed. Contrarily, inhibition of protease by its potential inhibitor decreased the voltammetric response. To demonstrate the feasibility and sensitivity of our strategy, beta-secretase was tested as a model protease. As a result, a detection limit of 0.08 nmol L-1 was achieved. The half-maximum inhibition value (IC50) of a well-known beta-secretase inhibitor was found to be 22.6 nmol L-1, which is in agreement with the reported value. (C) 2016 Elsevier B.V. All rights reserved.
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