Detection for folding of the thrombin binding aptamer using label-free electrochemical methods

被引:14
作者
Cho, Minseon [1 ]
Kim, Yeonwha [1 ]
Han, Se-Young [1 ]
Min, Kyungin [1 ]
Rahman, Aminur [2 ]
Shim, Yoon-Bo [2 ]
Ban, Changill [1 ]
机构
[1] Pohang Univ Sci & Technol, Dept Chem, Pohang, South Korea
[2] Pusan Natl Univ, Dept Chem, Pusan, South Korea
关键词
aptamer folding; impedance; thrombin; thrombin aptamer; XPS;
D O I
10.5483/BMBRep.2008.41.2.126
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The folding of aptamer immobilized on an Au electrode was successfully detected using label-free electrochemical methods. A thrombin binding DNA aptamer was used as a model system in the presence of various monovalent cations. Impedance spectra showed that the extent to which monovalent cations assist in folding of aptamer is ordered as K+ > NH4+ > Na+ > Cs+. Our XPS analysis also showed that K+ and NH4+ caused a conformational change of the aptamer in which it forms a stable complex with these monovalent ions. Impedance results for the interaction between aptamer and thrombin indicated that thrombin interacts more with folded aptamer than with unfolded aptamer. The EQCM technique provided a quantitative analysis of these results. In particular, the present impedance results showed that thrombin participates a folding of aptamer to some extent and XPS analysis confirmed that thrombin stabilizes and induces the folding of aptamer.
引用
收藏
页码:126 / 131
页数:6
相关论文
共 30 条
[1]  
BRIGGS D, 1996, PRACTICAL SURFACE AN, P112
[2]   Electrochemical detection of mismatched DNA using a MutS probe [J].
Cho, Minseon ;
Lee, Sohyun ;
Han, Se-Young ;
Park, Jin-Young ;
Rahman, Md Aminur ;
Shim, Yoon-Bo ;
Ban, Changill .
NUCLEIC ACIDS RESEARCH, 2006, 34 (10)
[3]   Development of an immunosensor for the detection of vitellogenin using impedance spectroscopy [J].
Darain, F ;
Park, DS ;
Park, JS ;
Shim, YB .
BIOSENSORS & BIOELECTRONICS, 2004, 19 (10) :1245-1252
[4]   Development of a new and simple method for the detection of histidine-tagged proteins [J].
Darain, F ;
Ban, CI ;
Shim, YB .
BIOSENSORS & BIOELECTRONICS, 2004, 20 (04) :857-863
[5]   Molecular aptamer for real-time oncoprotein platelet-derived growth factor monitoring by fluorescence anisotropy [J].
Fang, XH ;
Cao, ZH ;
Beck, T ;
Tan, WH .
ANALYTICAL CHEMISTRY, 2001, 73 (23) :5752-5757
[6]   Telomeres [J].
Greider, Carol W. .
CURRENT OPINION IN CELL BIOLOGY, 1991, 3 (03) :444-451
[7]   Aptamer beacons for the direct detection of proteins [J].
Hamaguchi, N ;
Ellington, A ;
Stanton, M .
ANALYTICAL BIOCHEMISTRY, 2001, 294 (02) :126-131
[8]   Quantum-dot/aptamer-based ultrasensitive multi-analyte electrochemical biosensor [J].
Hansen, JA ;
Wang, J ;
Kawde, AN ;
Xiang, Y ;
Gothelf, KV ;
Collins, G .
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 2006, 128 (07) :2228-2229
[9]   Biochemistry - Adaptive recognition by nucleic acid aptamers [J].
Hermann, T ;
Patel, DJ .
SCIENCE, 2000, 287 (5454) :820-825
[10]   Detection of aptamer-protein interactions using QCM and electrochemical indicator methods [J].
Hianik, T ;
Ostatná, V ;
Zajacová, Z ;
Stoikova, E ;
Evtugyn, G .
BIOORGANIC & MEDICINAL CHEMISTRY LETTERS, 2005, 15 (02) :291-295