D'Auria S, Di Cesare N, Gryczynski Z, Gryczynski I, Rossi M, Lakowicz J R
Department of Biochemistry and Molecular Biology, Center for Fluorescence Spectroscopy, University of Maryland at Baltimore, 725 West Lombard Street, Baltimore, Maryland, 21201, USA. dafne.ibpe.na.cnr.it
Biochem Biophys Res Commun. 2000 Aug 11;274(3):727-31. doi: 10.1006/bbrc.2000.3172.
Blood glucose is a clinically important analytes for diabetic health care. In this preliminary report we describe a protein biosensor for d-glucose based on a thermostable glucose dehydrogenase. The glucose dehydrogenase was noncovalently labeled with 8-anilino-1-naphthalene sulfonic acid (ANS). The ANS-labeled enzyme displayed an approximate 25% decrease in emission intensity upon binding glucose. This decrease can be used to measure the glucose concentration. Our results suggest that enzymes which use glucose as their substrate can be used as reversible and nonconsuming glucose sensors in the absence of required cofactors. Moreover, the possibility of using inactive apoenzymes for a reversible sensor greatly expands the range of proteins which can be used as sensors, not only for glucose, but for a wide variety of biochemically relevant analytes.
血糖是糖尿病医疗保健中一项具有临床重要意义的分析物。在本初步报告中,我们描述了一种基于热稳定葡萄糖脱氢酶的d-葡萄糖蛋白质生物传感器。葡萄糖脱氢酶用8-苯胺基-1-萘磺酸(ANS)进行非共价标记。结合葡萄糖后,ANS标记的酶发射强度降低了约25%。这种降低可用于测量葡萄糖浓度。我们的结果表明,以葡萄糖为底物的酶在缺乏所需辅因子的情况下可作为可逆且不消耗的葡萄糖传感器。此外,将无活性的脱辅基酶用于可逆传感器的可能性极大地扩展了可用作传感器的蛋白质范围,不仅可用于葡萄糖,还可用于多种与生物化学相关的分析物。