title: Novel cell-penetrating peptide-adaptors effect intracellular delivery and endosomal escape of protein cargos.
abstract: The use of cell-penetrating peptides (CPPs) as biomolecular delivery vehicles holds great promise for therapeutic and other applications, but development has been stymied by poor delivery and lack of endosomal escape. We have developed a CPP-adaptor system capable of efficient intracellular delivery and endosomal escape of user-defined protein cargos. The cell-penetrating sequence of HIV transactivator of transcription was fused to calmodulin, which binds with subnanomolar affinity to proteins containing a calmodulin binding site. Our strategy has tremendous advantage over prior CPP technologies because it utilizes high-affinity non-covalent, but reversible coupling between CPP and cargo. Three different cargo proteins fused to a calmodulin binding sequence were delivered to the cytoplasm of eukaryotic cells and released, demonstrating the feasibility of numerous applications in living cells including alteration of signaling pathways and gene expression.
authors.affiliation: Department of Molecular & Cellular Biology and Kennesaw State University, Kennesaw, GA 30144, USA.
authors.forename: John C
authors.initials: JC
authors.lastname: Salerno
authors.affiliation: Department of Chemistry & Biochemistry, Kennesaw State University, Kennesaw, GA 30144, USA.
authors.forename: Verra M
authors.initials: VM
authors.lastname: Ngwa
authors.affiliation: Department of Molecular & Cellular Biology and Kennesaw State University, Kennesaw, GA 30144, USA.
authors.forename: Scott J
authors.initials: SJ
authors.lastname: Nowak
authors.affiliation: Department of Chemistry & Biochemistry, Kennesaw State University, Kennesaw, GA 30144, USA.
authors.forename: Carol A
authors.initials: CA
authors.lastname: Chrestensen
authors.affiliation: New Echota Biotechnology, Kennesaw, GA 30144, USA.
authors.forename: Allison N
authors.initials: AN
authors.lastname: Healey
authors.affiliation: Department of Molecular & Cellular Biology and Kennesaw State University, Kennesaw, GA 30144, USA jmcmurr1@kennesaw.edu.
authors.forename: Jonathan L
authors.initials: JL
authors.lastname: McMurry
mesh_terms: D002147:Calmodulin / Q000737:chemistry; D057846:Cell-Penetrating Peptides / Q000737:chemistry / Q000378:metabolism*; D011992:Endosomes / Q000378:metabolism*; D015696:Gene Products, tat / Q000737:chemistry; D057809:HEK293 Cells; D006801:Humans; D009211:Myoglobin / Q000378:metabolism*; D021381:Protein Transport; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism*
chemical_list: D002147:Calmodulin; D057846:Cell-Penetrating Peptides; D015696:Gene Products, tat; D009211:Myoglobin; D011993:Recombinant Fusion Proteins
keywords: Cell-penetrating peptides; Cellular delivery; Confocal microscopy; Protein transduction domains; Protein transport; TAT