title: Channel of viral DNA packaging motor for real time kinetic analysis of peptide oxidation states.
abstract: Nanopore technology has become a powerful tool in single molecule sensing, and protein nanopores appear to be more advantageous than synthetic counterparts with regards to channel amenability, structure homogeneity, and production reproducibility. However, the diameter of most of the well-studied protein nanopores is too small to allow the passage of protein or peptides that are typically in multiple nanometers scale. The portal channel from bacteriophage SPP1 has a large channel size that allows the translocation of peptides with higher ordered structures. Utilizing single channel conductance assay and optical single molecule imaging, we observed translocation of peptides and quantitatively analyzed the dynamics of peptide oligomeric states in real-time at single molecule level. The oxidative and the reduced states of peptides were clearly differentiated based on their characteristic electronic signatures. A similar Gibbs free energy (ΔG0) was obtained when different concentrations of substrates were applied, suggesting that the use of SPP1 nanopore for real-time quantification of peptide oligomeric states is feasible. With the intrinsic nature of size and conjugation amenability, the SPP1 nanopore has the potential for development into a tool for the quantification of peptide and protein structures in real time.
authors.affiliation: College of Pharmacy, Division of Pharmaceutics and Pharmaceutical Chemistry; College of Medicine, Department of Physiology & Cell Biology; and Dorothy M. Davis Heart and Lung Research Institute, The Ohio State University, Columbus, Ohio 43210, USA; College of Pharmacy, Markey Cancer Center, University of Kentucky, Lexington, KY 40536, USA.
authors.forename: Shaoying
authors.initials: S
authors.lastname: Wang
authors.affiliation: College of Pharmacy, Division of Pharmaceutics and Pharmaceutical Chemistry; College of Medicine, Department of Physiology & Cell Biology; and Dorothy M. Davis Heart and Lung Research Institute, The Ohio State University, Columbus, Ohio 43210, USA.
authors.forename: Zhi
authors.initials: Z
authors.lastname: Zhou
authors.affiliation: College of Pharmacy, Division of Pharmaceutics and Pharmaceutical Chemistry; College of Medicine, Department of Physiology & Cell Biology; and Dorothy M. Davis Heart and Lung Research Institute, The Ohio State University, Columbus, Ohio 43210, USA; College of Pharmacy, Markey Cancer Center, University of Kentucky, Lexington, KY 40536, USA.
authors.forename: Zhengyi
authors.initials: Z
authors.lastname: Zhao
authors.affiliation: College of Pharmacy, Division of Pharmaceutics and Pharmaceutical Chemistry; College of Medicine, Department of Physiology & Cell Biology; and Dorothy M. Davis Heart and Lung Research Institute, The Ohio State University, Columbus, Ohio 43210, USA.
authors.forename: Hui
authors.initials: H
authors.lastname: Zhang
authors.affiliation: College of Pharmacy, Division of Pharmaceutics and Pharmaceutical Chemistry; College of Medicine, Department of Physiology & Cell Biology; and Dorothy M. Davis Heart and Lung Research Institute, The Ohio State University, Columbus, Ohio 43210, USA.
authors.forename: Farzin
authors.initials: F
authors.lastname: Haque
authors.affiliation: College of Pharmacy, Division of Pharmaceutics and Pharmaceutical Chemistry; College of Medicine, Department of Physiology & Cell Biology; and Dorothy M. Davis Heart and Lung Research Institute, The Ohio State University, Columbus, Ohio 43210, USA. Electronic address: guo.1091@osu.edu.
authors.forename: Peixuan
authors.initials: P
authors.lastname: Guo
mesh_terms: D001435:Bacteriophages; D042003:DNA Packaging; D004279:DNA, Viral / Q000737:chemistry*; D007700:Kinetics; D008051:Lipid Bilayers / Q000737:chemistry; D008856:Microscopy, Fluorescence; D058608:Nanopores; D010084:Oxidation-Reduction; D010455:Peptides / Q000737:chemistry*; D011487:Protein Conformation; D015203:Reproducibility of Results; D054322:tat Gene Products, Human Immunodeficiency Virus / Q000378:metabolism
chemical_list: D004279:DNA, Viral; D008051:Lipid Bilayers; D010455:Peptides; D054322:tat Gene Products, Human Immunodeficiency Virus
keywords: Bacteriophage assembly; Biomotor; Nanobiotechnology; Nanopore sensing; Peptide identification; Viral motor