title: Engineering PQQ-glucose dehydrogenase into an allosteric electrochemical Ca(2+) sensor.
abstract: 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 Ca(2+) biosensor that could be used to rapidly measure Ca(2+) concentrations in human biological fluids. We were able to convert a point-of-care glucometer into Ca(2+) 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.
authors.affiliation: Institute for Molecular Bioscience, The University of Queensland, Brisbane, QLD 4072, Australia. k.alexandrov@uq.edu.au.
authors.forename: Zhong
authors.initials: Z
authors.lastname: Guo
authors.forename: Wayne A
authors.initials: WA
authors.lastname: Johnston
authors.forename: Viktor
authors.initials: V
authors.lastname: Stein
authors.forename: Palraj
authors.initials: P
authors.lastname: Kalimuthu
authors.forename: Siro
authors.initials: S
authors.lastname: Perez-Alcala
authors.forename: Paul V
authors.initials: PV
authors.lastname: Bernhardt
authors.forename: Kirill
authors.initials: K
authors.lastname: Alexandrov
mesh_terms: D000494:Allosteric Regulation; D015374:Biosensing Techniques; D002118:Calcium / Q000737:chemistry*; D002147:Calmodulin / Q000737:chemistry; D055664:Electrochemical Techniques; D042843:Glucose 1-Dehydrogenase / Q000737:chemistry*; D006801:Humans; D015202:Protein Engineering
chemical_list: D002147:Calmodulin; D042843:Glucose 1-Dehydrogenase; D002118:Calcium