title: Systematic screening of the cellular uptake of designed alpha-helix peptides.
abstract: The cellular penetration (CP) activity of functional molecules has attracted significant attention as one of the most promising new approaches for drug delivery. In particular, cell-penetrating peptides (CPPs) have been studied extensively in cellular engineering. Because there have been few large-scale systematic studies to identify peptide sequences with optimal CP activity or that are suitable for further applications in cell engineering, such as cell-specific penetration and cell-selective culture, we screened and compared the cellular uptake (CU) activity of 54 systematically designed α-helical peptides in HeLa cells. Furthermore, the CU activity of 24 designed peptides was examined in four cell lines using a cell fingerprinting technique and statistical approaches. The CU activities in various cells depended on amino acid residues of peptide sequences as well as charge, α-helical content and hydrophobicity of the peptides. Notably, the mutation of a single residue significantly altered the CU ability of a peptide, highlighting the variability of cell uptake mechanisms. Moreover, these results demonstrated the feasibility of cell-selective culture by conducting cell-selective permeation and death in cultures containing two cell types. These studies may lead to further peptide library design and screening for new classes of CPPs with useful functions.
authors.affiliation: Department of Bioengineering, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, Yokohama 226-8501, Japan. kusui@center.konan-u.ac.jp
authors.forename: Kenji
authors.initials: K
authors.lastname: Usui
authors.forename: Takuya
authors.initials: T
authors.lastname: Kikuchi
authors.forename: Masayasu
authors.initials: M
authors.lastname: Mie
authors.forename: Eiry
authors.initials: E
authors.lastname: Kobatake
authors.forename: Hisakazu
authors.initials: H
authors.lastname: Mihara
mesh_terms: D041721:3T3-L1 Cells; D000818:Animals; D002470:Cell Survival; D002478:Cells, Cultured; D015195:Drug Design; D006367:HeLa Cells; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D051379:Mice; D016716:PC12 Cells; D019151:Peptide Library; D010455:Peptides / Q000138:chemical synthesis / Q000737:chemistry / Q000493:pharmacokinetics*; D017433:Protein Structure, Secondary; D051381:Rats
chemical_list: D019151:Peptide Library; D010455:Peptides