title: Fingerprinting of Peptides with a Large Channel of Bacteriophage Phi29 DNA Packaging Motor.
abstract: Nanopore technology has become a highly sensitive and powerful tool for single molecule sensing of chemicals and biopolymers. Protein pores have the advantages of size amenability, channel homogeneity, and fabrication reproducibility. But most well-studied protein pores for sensing are too small for passage of peptide analytes that are typically a few nanometers in dimension. The funnel-shaped channel of bacteriophage phi29 DNA packaging motor has previously been inserted into a lipid membrane to serve as a larger pore with a narrowest N-terminal constriction of 3.6 nm and a wider C-terminal end of 6 nm. Here, the utility of phi29 motor channel for fingerprinting of various peptides using single molecule electrophysiological assays is reported. The translocation of peptides is proved unequivocally by single molecule fluorescence imaging. Current blockage percentage and distinctive current signatures are used to distinguish peptides with high confidence. Each peptide generated one or two distinct current blockage peaks, serving as typical fingerprint for each peptide. The oligomeric states of peptides can also be studied in real time at single molecule level. The results demonstrate the potential for further development of phi29 motor channel for detection of disease-associated peptide biomarkers.
authors.affiliation: College of Pharmacy, Division of Pharmaceutics and Pharmaceutical Chemistry, College of Medicine, Department of Physiology and Cell Biology, Dorothy M. Davis Heart and Lung Research Institute, The Ohio State University, Columbus, OH, 43210, USA.
authors.forename: Zhouxiang
authors.initials: Z
authors.lastname: Ji
authors.affiliation: College of Pharmacy, Division of Pharmaceutics and Pharmaceutical Chemistry, College of Medicine, Department of Physiology and Cell Biology, Dorothy M. Davis Heart and Lung Research Institute, The Ohio State University, Columbus, OH, 43210, 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 and Cell Biology, Dorothy M. Davis Heart and Lung Research Institute, The Ohio State University, Columbus, OH, 43210, 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 and Cell Biology, Dorothy M. Davis Heart and Lung Research Institute, The Ohio State University, Columbus, OH, 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 and Cell Biology, Dorothy M. Davis Heart and Lung Research Institute, The Ohio State University, Columbus, OH, 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 and Cell Biology, Dorothy M. Davis Heart and Lung Research Institute, The Ohio State University, Columbus, OH, 43210, USA.
authors.forename: Peixuan
authors.initials: P
authors.lastname: Guo
mesh_terms: D001435:Bacteriophages / Q000737:chemistry*; D042003:DNA Packaging; D005453:Fluorescence; D008051:Lipid Bilayers / Q000737:chemistry; D010449:Peptide Mapping / Q000379:methods*; D010455:Peptides / Q000737:chemistry*; D015203:Reproducibility of Results; D013997:Time Factors
chemical_list: D008051:Lipid Bilayers; D010455:Peptides
keywords: nanopores; nanotechnology; phi29 connector; single molecule sensing