A FRET-based biosensor for NO detection

被引：19
|
作者：
Strianese, Maria [2 ]
De Martino, Franco [2 ]
Pavone, Vincenzo [3 ]
Lombardi, Angela [3 ]
Canters, Gerard W. [1 ]
Pellecchia, Claudio [2 ]
机构：
[1] Leiden Univ, Leiden Inst Chem, NL-2333 CC Leiden, Netherlands
[2] Univ Salerno, Dipartimento Chim, I-84084 Fisciano, Sa, Italy
[3] Univ Naples Federico II, Dipartimento Chim, I-80126 Naples, Italy
来源：
JOURNAL OF INORGANIC BIOCHEMISTRY | 2010年 / 104卷 / 06期
关键词：
Heme proteins; Biosensors; Fluorescence; FRET; Sol-gel processes; CYTOCHROME-C PEROXIDASE; NITRIC-OXIDE CONCENTRATIONS; ENERGY-TRANSFER; CELLULAR APPLICATIONS; TUMOR PROGRESSION; BINDING; YEAST; LIPOPOLYSACCHARIDE; COORDINATION; GENERATION;
D O I：
10.1016/j.jinorgbio.2010.02.002
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：
In this paper we explore the use of fluorescently labeled cytochrome c peroxidase (CcP) from baker's yeast for monitoring nitric oxide (NO) down to the sub-micromolar level, by means of a FRET (Forster Resonance Energy Transfer) mechanism. The binding affinity constant (K-d) for the NO binding to CcP was determined to be 10 +/- 1.5 mu M. The rate of NO dissociation from the CcP (k(off)) and the second order rate constant for the NO association (k(on)) were found to be 0.22 +/- 0.08 min(-1) and 0.024 +/- 0.002 mu M-1 min(-1) respectively. The immobilization of fluorescently labeled CcP into a polymeric matrix for use in a solid state NO sensing device was also explored. The results provide proof-of-principle that labeled CcP can be successfully implemented in a fast, simple, quantitative and sensitive NO sensing device. (C) 2010 Elsevier Inc. All rights reserved.
引用
收藏
页码：619 / 624
页数：6
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