Engineering PQQ-glucose dehydrogenase into an allosteric electrochemical Ca2+ sensor

被引:33
|
作者
Guo, Zhong [1 ]
Johnston, Wayne A. [1 ]
Stein, Viktor [1 ]
Kalimuthu, Palraj [2 ]
Perez-Alcala, Siro [3 ]
Bernhardt, Paul V. [2 ]
Alexandrov, Kirill [1 ]
机构
[1] Univ Queensland, Inst Mol Biosci, Brisbane, Qld 4072, Australia
[2] Univ Queensland, Sch Chem & Mol Biosci, Brisbane, Qld 4072, Australia
[3] Mol Warehouse Ltd, Twickenham TW1 3DY, England
基金
澳大利亚研究理事会; 澳大利亚国家健康与医学研究理事会;
关键词
PYRROLOQUINOLINE-QUINONE; ACINETOBACTER-CALCOACETICUS; PROTEIN SWITCHES; CALMODULIN; CALCIUM; RECONSTITUTION; INDICATORS; BIOSENSORS; DESIGN;
D O I
10.1039/c5cc07824e
中图分类号
O6 [化学];
学科分类号
0703 ;
摘要
Electrochemical biosensors convert biological events to an electrical current. To date most electrochemical biosensors exploit activities of naturally occurring enzymes. Here we demonstrated that insertion of a calmodulin domain into the redox enzyme PQQ-glucose dehydrogenase resulted in a selective Ca2+ biosensor that could be used to rapidly measure Ca2+ concentrations in human biological fluids. We were able to convert a point-of-care glucometer into Ca2+ monitor by refurbishing it with the developed biosensor. We propose that similar engineering strategies may be used to create highly specific electrochemical biosensors to other analytes. Compatibility with cheap and ubiquitous amperometric detectors is expected to accelerate progression of these biosensors into clinical applications.
引用
收藏
页码:485 / 488
页数:4
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