title: Using the peptide BP100 as a cell-penetrating tool for the chemical engineering of actin filaments within living plant cells.
abstract: The delivery of externally applied macromolecules or nanoparticles into living cells still represents a critically limiting step before the full capabilities of chemical engineering can be explored. Molecular transporters such as cell-penetrating peptides, peptoids, and other mimetics can be used to carry cargo across the cellular membrane, but it is still difficult to find suitable sequences that operate efficiently for any particular type of cell. Here we report that BP100 (KKLFKKILKYL-amide), originally designed as an antimicrobial peptide against plant pathogens, can be employed as a fast and efficient cell-penetrating agent to transport fluorescent test cargoes into the cytosol of walled plant cells. The uptake of BP100 proceeds slightly more slowly than the endocytosis of fluorescent dextranes, but BP100 accumulates more efficiently and to much higher levels (by an order of magnitude). The entry of BP100 can be efficiently blocked by latrunculin B; this suggests that actin filaments are essential to the uptake mechanism. To test whether this novel transporter can also be used to deliver functional cargoes, we designed a fusion construct of BP100 with the actin-binding Lifeact peptide (MGVADLIKKFESISKEE). We demonstrated that the short BP100 could transport the attached 17-residue sequence quickly and efficiently into tobacco cells. The Lifeact construct retained its functionality as it successfully labeled the actin bundles that tether the nucleus in the cell center.
authors.affiliation: Molecular Cell Biology, Institute of Botany, DFG-Center of Functional Nanostructures (CFN), Karlsruhe Institute of Technology (KIT), Kaiserstrasse 2, 76131 Karlsruhe, Germany. kai.eggenberger@kit.edu
authors.forename: Kai
authors.initials: K
authors.lastname: Eggenberger
authors.forename: Christian
authors.initials: C
authors.lastname: Mink
authors.forename: Parvesh
authors.initials: P
authors.lastname: Wadhwani
authors.forename: Anne S
authors.initials: AS
authors.lastname: Ulrich
authors.forename: Peter
authors.initials: P
authors.lastname: Nick
mesh_terms: D008841:Actin Cytoskeleton / Q000378:metabolism*; D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D002460:Cell Line; D002470:Cell Survival; D057846:Cell-Penetrating Peptides / Q000737:chemistry* / Q000378:metabolism*; D002616:Chemical Engineering / Q000379:methods*; D003593:Cytoplasm / Q000378:metabolism; D004705:Endocytosis; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000737:chemistry* / Q000378:metabolism*; D021381:Protein Transport; D014026:Nicotiana / Q000166:cytology* / Q000378:metabolism
chemical_list: D057846:Cell-Penetrating Peptides; D009842:Oligopeptides; C540525:lysyl-lysyl-leucyl-phenylalanyl-lysyl-lysyl-isoleucyl-leucyl-lysyl-tyrosyl-leucinamide