title: Bioluminescence of monolayers of firefly luciferase immobilized on graphite.
abstract: We report on the immobilization of the firefly protein luciferase on the hydrophobic surface of graphite. Observation by liquid-phase atomic force microscopy of islands with a height consistent with the size of a single molecule confirmed that the protein was contained within a monomolecular layer. The enzyme activity was assayed by single-photon counting of the bioluminescence, which is the catalytic product of luciferase. Attachment to the surface modified the efficiency of the enzyme, but the introduction of the substrates luciferin and ATP resulted in the reactivation of the enzyme. The functionalized graphite surface was employed as a cathode in a bioelectrochemical cell. This demonstrated that the electric field caused a substantial loss of enzyme catalytic activity.
authors.affiliation: Nanoscale Physics Research Laboratory, School of Physics and Astronomy, University of Birmingham, Edgbaston, Birmingham B15 2TT, UK.
authors.forename: S
authors.initials: S
authors.lastname: Palomba
authors.forename: N
authors.initials: N
authors.lastname: Berovic
authors.forename: R E
authors.initials: RE
authors.lastname: Palmer
mesh_terms: D000255:Adenosine Triphosphate / Q000737:chemistry*; D004563:Electrochemistry; D004789:Enzyme Activation; D004800:Enzymes, Immobilized / Q000737:chemistry; D049454:Firefly Luciferin / Q000737:chemistry*; D006108:Graphite / Q000737:chemistry*; D049409:Luciferases, Firefly / Q000737:chemistry*; D049449:Luminescence
chemical_list: D004800:Enzymes, Immobilized; D049454:Firefly Luciferin; D006108:Graphite; D000255:Adenosine Triphosphate; D049409:Luciferases, Firefly