Glucose sensor for low-cost lifetime-based sensing using a genetically engineered protein
Glucose sensor for low-cost lifetime-based sensing using a genetically engineered protein
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DOI：
10.1006/abio.1998.2974
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发表时间：
1999-02-01
影响因子：
2.9
通讯作者：
Lakowicz, JR
中科院分区：
文献类型：
--
作者：
Tolosa, L;Gryczynski, I;Lakowicz, JR
我们描述了一种基于突变型葡萄糖/半乳糖结合蛋白（GGBP）和相位调制荧光法的葡萄糖传感器。来自大肠杆菌的GGBP突变为在位置26处含有单个半胱氨酸残基。当用巯基反应性探针2-（4 '-iodoacetamidanilino）naphthalene-6-sulfonic acid标记时，标记的蛋白质对葡萄糖的反应强度降低两倍，解离常数接近1 μ M葡萄糖。ANS标记的蛋白质在寿命上仅显示出适度的变化，排除了基于寿命的葡萄糖感测。通过将ANS 26-GGBP与比色皿表面上的长寿命钌（Ru）金属配体络合物结合来创建调制传感器。葡萄糖的结合改变了ANS 26-GGBP和Ru络合物的相对强度，导致在2.1 MHz的低频下调制的急剧变化。在2.1 MHz的调制测量显示，以准确地确定葡萄糖浓度。这些结果提出了一种利用简单设备进行葡萄糖感测的方法。(C)北京：科学出版社.
We describe a glucose sensor based on a mutant glucose/galactose binding protein (GGBP) and phase-modulation fluorometry. The GGBP from Escherichia coli was mutated to contain a single cysteine residue at position 26. When labeled with a sulfhydryl-reactive probe 2-(4'-iodoacetamidoanilino)naphthalene-6-sulfonic acid, the labeled protein displayed a twofold decrease in intensity in response to glucose, with a dissociation constant near 1 mu M glucose. The ANS-labeled protein displayed only a modest change in lifetime, precluding lifetime-based sensing of glucose. A modulation sensor was created by combining ANS26-GGBP with a long-lifetime ruthenium (Ru) metal-ligand complex on the surface of the cuvette. Binding of glucose changed the relative intensity of ANS26-GGBP and the Ru complex, resulting in a dramatic change in modulation at a low frequency of 2.1 MHz. Modulation measurements at 2.1 MHz were shown to accurately determine the glucose concentration. These results suggest an approach to glucose sensing with simple devices. (C) 1999 Academic Press.