Photoswitchable peptide-based ‘on-off’ biosensor for electrochemical detection and control of protein-protein interactions

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dc.contributor.authorHorsley, J.R.
dc.contributor.authorYu, J.
dc.contributor.authorWegener, K.L.
dc.contributor.authorHoppmann, C.
dc.contributor.authorRück-Braun, K.
dc.contributor.authorAbell, A.D.
dc.date.issued2018
dc.description.abstractNeuronal nitric oxide synthase (nNOS) is an enzyme responsible for catalyzing the production of the crucial cellular signalling molecule, nitric oxide (NO), through its interaction with the PDZ domain of α-syntrophin protein. In this study, a novel light-driven photoswitchable peptide-based biosensor, modelled on the nNOS β-finger, is used to detect and control its interaction with α-syntrophin. An azobenzene photoswitch incorporated into the peptide backbone allows reversible switching between a trans photostationary state devoid of secondary structure, and a cis photostationary state possessing a well-defined antiparallel β-strand geometry, as revealed by molecular modelling. Electrochemical impedance spectroscopy (EIS) is used to successfully detect the interaction between the gold electrode bound peptide in its cis photostationary state and a wide range of concentrations of α-syntrophin protein, highlighting both the qualitative and quantitative properties of the sensor. Furthermore, EIS demonstrates that the probe in its random trans photostationary state does not bind to the target protein. The effectiveness of the biosensor is further endorsed by the high thermal stability of the photostationary state of the cis-isomer, and the ability to actively control biomolecular interactions using light. This approach allows detection and control of binding to yield a regenerable on-off biosensor.
dc.description.statementofresponsibilityJohn R.Horsley, Jingxian Yu, Kate L.Wegener, Christian Hoppmann, Karola Rück-Braun, Andrew D.Abell
dc.identifier.citationBiosensors and Bioelectronics, 2018; 118:188-194
dc.identifier.doi10.1016/j.bios.2018.07.057
dc.identifier.issn0956-5663
dc.identifier.issn1873-4235
dc.identifier.orcidYu, J. [0000-0002-6489-5988]
dc.identifier.orcidWegener, K.L. [0000-0002-1562-6060]
dc.identifier.orcidAbell, A.D. [0000-0002-0604-2629]
dc.identifier.urihttp://hdl.handle.net/2440/116619
dc.language.isoen
dc.publisherElsevier
dc.relation.granthttp://purl.org/au-research/grants/arc/CE140100003
dc.relation.granthttp://purl.org/au-research/grants/nhmrc/1068087
dc.rights© 2018 Elsevier B.V. All rights reserved.
dc.source.urihttps://doi.org/10.1016/j.bios.2018.07.057
dc.subjectBiosensors
dc.subjectElectrochemical impedance spectroscopy
dc.subjectNeuronal nitric oxide synthase (nNOS)
dc.subjectPhotoswitchable peptides
dc.subjectProtein-protein interaction
dc.subjectSecondary structure
dc.titlePhotoswitchable peptide-based ‘on-off’ biosensor for electrochemical detection and control of protein-protein interactions
dc.typeJournal article
pubs.publication-statusPublished

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