title: Direct electrochemistry of novel affinity-tag immobilized recombinant horse heart cytochrome c.
abstract: During the last decade protein electrochemistry at miniaturized electrodes has become important not only for functional studies of the charge transfer properties of redox proteins but also for fostering the development of sensitive biosensor and bioelectronic devices. One of the major challenges in this field is the directed coupling between electronic and biologically active components. A prerequisite for a fast and reversible electron transfer between electrode and protein is that the protein can be bound to the electrode in a favourable orientation. We examined electrostatic and bioaffinity-tag binding strategies for the directed immobilization of horse heart cytochrome c (cytc) on gold electrode surfaces to achieve this goal. Horse heart cytc was expressed in E. coli either as non-modified or genetically modified, i.e. histidine (his)-tag containing protein. The his-tags were introduced at defined positions at the N- or C-terminus of the polypeptide. It was our aim to generate tagged-versions of cytc that facilitate strong electronic coupling between protein and electrode and, at the same time, retain their catalytic and regulatory properties. The combination of different immobilization strategies, e.g. his-tag and electrostatic immobilization also opens new avenues for bivalent immobilization of proteins. This is of interest for molecular bioelectronic and biosensing applications where the proteins are immobilized between two crossing electrodes.
authors.affiliation: Peter Grünberg Institute (PGI-8), Forschungszentrum Jülich, Jülich, Germany.
authors.forename: Florian
authors.initials: F
authors.lastname: Schröper
authors.forename: Arnd
authors.initials: A
authors.lastname: Baumann
authors.forename: Andreas
authors.initials: A
authors.lastname: Offenhäusser
authors.forename: Dirk
authors.initials: D
authors.lastname: Mayer
mesh_terms: D000818:Animals; D015374:Biosensing Techniques / Q000379:methods*; D045304:Cytochromes c / Q000737:chemistry* / Q000235:genetics; D055664:Electrochemical Techniques / Q000379:methods*; D004566:Electrodes; D004579:Electron Transport; D004800:Enzymes, Immobilized / Q000737:chemistry* / Q000235:genetics; D006046:Gold / Q000737:chemistry; D006321:Heart; D006736:Horses; D011994:Recombinant Proteins / Q000737:chemistry* / Q000235:genetics
chemical_list: D004800:Enzymes, Immobilized; D011994:Recombinant Proteins; D006046:Gold; D045304:Cytochromes c