title: Identification and evaluation of a novel peptide binding to the cell surface of Staphylococcus aureus.
abstract: Identification of short peptides that serve as specific ligands to biological materials such as microbial cell surfaces has major implications in better understanding the molecular recognition of cell surfaces. In this study we screened a commercially available random phage-display library against Staphylococcus aureus cells and identified peptides specifically binding to the bacteria. A synthetic peptide (SA5-1) representing the consensus sequence (VPHNPGLISLQG) of the bacteria-binding peptide was evaluated for its binding potential against S. aureus. Dot-blot, immunoblot assay and ELISA results revealed the SA5-1 peptide to be highly specific to S. aureus. The SA5-1 peptide binding was optimal between pH 6.0 and 8.0. Nanogold Transmission Electron Microscopy demonstrated that the SA5-1 binds to the outer membrane surface of S. aureus. Diagnostic potential of the SA5-1 peptide was evaluated in human platelet samples spiked with S. aureus and specific detection of the bacteria by biotinylated-SA5-1 and streptavidin-conjugated fluorescent quantum dots. Fluorometry results indicated that the peptide was able to detect ∼100 organisms per ml in a spiked biological sample providing a proof-of-concept towards potential of this peptide as a S. aureus diagnostic tool that can be of use in different detection platforms.
authors.affiliation: Section of Cell Biology, Laboratory of Cellular Hematology, Division of Hematology, Center for Biologics Evaluation and Research, FDA, Bethesda, MD 20892, USA.
authors.forename: Shilpakala Sainath
authors.initials: SS
authors.lastname: Rao
authors.forename: Ketha V Krishna
authors.initials: KV
authors.lastname: Mohan
authors.forename: Yamei
authors.initials: Y
authors.lastname: Gao
authors.forename: Chintamani D
authors.initials: CD
authors.lastname: Atreya
mesh_terms: D000595:Amino Acid Sequence; D002462:Cell Membrane / Q000378:metabolism* / Q000648:ultrastructure; D016384:Consensus Sequence; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D008024:Ligands; D046529:Microscopy, Electron, Transmission; D008969:Molecular Sequence Data; D019151:Peptide Library; D010455:Peptides / Q000737:chemistry* / Q000378:metabolism*; D011485:Protein Binding; D045663:Quantum Dots; D013045:Species Specificity; D013203:Staphylococcal Infections / Q000175:diagnosis / Q000382:microbiology; D013211:Staphylococcus aureus / Q000378:metabolism* / Q000648:ultrastructure*
chemical_list: D008024:Ligands; D019151:Peptide Library; D010455:Peptides