title: Single molecule recordings of lysozyme activity.
abstract: Single molecule bioelectronic circuits provide an opportunity to study chemical kinetics and kinetic variability with bond-by-bond resolution. To demonstrate this approach, we examined the catalytic activity of T4 lysozyme processing peptidoglycan substrates. Monitoring a single lysozyme molecule through changes in a circuit's conductance helped elucidate unexplored and previously invisible aspects of lysozyme's catalytic mechanism and demonstrated lysozyme to be a processive enzyme governed by 9 independent time constants. The variation of each time constant with pH or substrate crosslinking provided different insights into catalytic activity and dynamic disorder. Overall, ten lysozyme variants were synthesized and tested in single molecule circuits to dissect the transduction of chemical activity into electronic signals. Measurements show that a single amino acid with the appropriate properties is sufficient for good signal generation, proving that the single molecule circuit technique can be easily extended to other proteins.
authors.affiliation: Department of Physics and Astronomy, University of California at Irvine, Irvine, California 92697, USA.
authors.forename: Yongki
authors.initials: Y
authors.lastname: Choi
authors.forename: Gregory A
authors.initials: GA
authors.lastname: Weiss
authors.forename: Philip G
authors.initials: PG
authors.lastname: Collins
mesh_terms: D055162:Biocatalysis; D007700:Kinetics; D008958:Models, Molecular; D009113:Muramidase / Q000737:chemistry / Q000378:metabolism*
chemical_list: D009113:Muramidase