Abstract
In this report, we established a new electrochemical method for the detection of conformational changes in large, non-metalloproteins such as bovine serum albumin, using flow injection analysis coupled with hydrogen-terminated, boron-doped diamond electrodes. The oxidation current was used as a signal reporter in the monitoring of urea-induced BSA denaturation. In the denatured state at high urea concentrations, the electrochemical signal increased, and the amperometric responses for the oxidation potential at 1300 mV were consistent with the results of conventional methods of denaturation monitoring using fluorescence spectroscopy. The oxidation involved at least five redox-active species (cysteine, tryptophan, tyrosine, methionine, and disulfide bonds). Furthermore, the method also showed high sensitivity for quantitative analysis of protein. A linear dynamic in the concentration range 50-400 μg/mL (r 2 = 0.977) with a lower detection limit of 190 ng/mL was achieved for BSA. Direct electrochemical detection of conformation changes of proteins using BDD electrodes can be performed with advantages in terms of simplicity and sensitivity.
| Original language | English |
|---|---|
| Pages (from-to) | 5783-5787 |
| Number of pages | 5 |
| Journal | Analytical Chemistry |
| Volume | 80 |
| Issue number | 15 |
| DOIs | |
| Publication status | Published - 2008 Aug 1 |
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ASJC Scopus subject areas
- Analytical Chemistry
Cite this
Conformational change detection in nonmetal proteins by direct electrochemical oxidation using diamond electrodes. / Chiku, Masanobu; Nakamura, Jin; Fujishima, Akira; Einaga, Yasuaki.
In: Analytical Chemistry, Vol. 80, No. 15, 01.08.2008, p. 5783-5787.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Conformational change detection in nonmetal proteins by direct electrochemical oxidation using diamond electrodes
AU - Chiku, Masanobu
AU - Nakamura, Jin
AU - Fujishima, Akira
AU - Einaga, Yasuaki
PY - 2008/8/1
Y1 - 2008/8/1
N2 - In this report, we established a new electrochemical method for the detection of conformational changes in large, non-metalloproteins such as bovine serum albumin, using flow injection analysis coupled with hydrogen-terminated, boron-doped diamond electrodes. The oxidation current was used as a signal reporter in the monitoring of urea-induced BSA denaturation. In the denatured state at high urea concentrations, the electrochemical signal increased, and the amperometric responses for the oxidation potential at 1300 mV were consistent with the results of conventional methods of denaturation monitoring using fluorescence spectroscopy. The oxidation involved at least five redox-active species (cysteine, tryptophan, tyrosine, methionine, and disulfide bonds). Furthermore, the method also showed high sensitivity for quantitative analysis of protein. A linear dynamic in the concentration range 50-400 μg/mL (r 2 = 0.977) with a lower detection limit of 190 ng/mL was achieved for BSA. Direct electrochemical detection of conformation changes of proteins using BDD electrodes can be performed with advantages in terms of simplicity and sensitivity.
AB - In this report, we established a new electrochemical method for the detection of conformational changes in large, non-metalloproteins such as bovine serum albumin, using flow injection analysis coupled with hydrogen-terminated, boron-doped diamond electrodes. The oxidation current was used as a signal reporter in the monitoring of urea-induced BSA denaturation. In the denatured state at high urea concentrations, the electrochemical signal increased, and the amperometric responses for the oxidation potential at 1300 mV were consistent with the results of conventional methods of denaturation monitoring using fluorescence spectroscopy. The oxidation involved at least five redox-active species (cysteine, tryptophan, tyrosine, methionine, and disulfide bonds). Furthermore, the method also showed high sensitivity for quantitative analysis of protein. A linear dynamic in the concentration range 50-400 μg/mL (r 2 = 0.977) with a lower detection limit of 190 ng/mL was achieved for BSA. Direct electrochemical detection of conformation changes of proteins using BDD electrodes can be performed with advantages in terms of simplicity and sensitivity.
UR - http://www.scopus.com/inward/record.url?scp=49449092317&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=49449092317&partnerID=8YFLogxK
U2 - 10.1021/ac800394n
DO - 10.1021/ac800394n
M3 - Article
C2 - 18613649
AN - SCOPUS:49449092317
VL - 80
SP - 5783
EP - 5787
JO - Analytical Chemistry
JF - Analytical Chemistry
SN - 0003-2700
IS - 15
ER -