title: Transduction of proteins into leishmania tarentolae by formation of non-covalent complexes with cell-penetrating peptides.
abstract: Cell-penetrating peptides (CPPs) are used to transport peptides, proteins, different types of ribonucleic acids (or mimics of these molecules), and DNA into live cells, both plant and mammalian. Leishmania belongs to the class of protozoa having, in comparison to mammalian cells, a different lipid composition of the membrane, proteoglycans on the surface, and signal pathways. We investigated the uptake of two different and easily detectable proteins into the non-pathogenic strain Leishmania tarentolae. From the large number of CPPs available, six and a histone were chosen specifically for their ability to form non-covalent complexes. For Leishmania we used the enzyme β-galactosidase and fluorescent labeled bovine serum albumin as cargoes. The results are compared to similar internalization studies using mammalian cells [Mussbach et al., ]. Leishmania cells can degrade CPPs by a secreted and membrane-bound chymotrypsin-like protease. Both cargo proteins were internalized with sufficient efficiency and achieved intramolecular concentrations similar to mammalian cells. The transport efficiencies of the CPPs differed from each other, and showed a different rank order for both cargoes. The intracellular distribution of fluorescent-labeled bovine serum albumin showed highest concentrations in the nucleus and kinetoplast. Leishmania are susceptible to high concentrations of some CPPs, although comparably dissimilar to mammalian cells. MPG-peptides are more cytotoxic in Leishmania than in mammalian cells, acting as antimicrobial peptides. Our results contribute to a better understanding of molecular interactions in Leishmania cells and possibly to new treatments of leishmaniasis.
authors.affiliation: Friedrich Schiller University, Biological and Pharmaceutical Faculty, Institute of Nutrition and Abbe Centre of Photonics, Dornburger Str. 25, 07743, Jena, Germany.
authors.forename: Andrea-Anneliese
authors.initials: AA
authors.lastname: Keller
authors.forename: Reinhard
authors.initials: R
authors.lastname: Breitling
authors.forename: Peter
authors.initials: P
authors.lastname: Hemmerich
authors.forename: Katarina
authors.initials: K
authors.lastname: Kappe
authors.forename: Maria
authors.initials: M
authors.lastname: Braun
authors.forename: Berith
authors.initials: B
authors.lastname: Wittig
authors.forename: Buerk
authors.initials: B
authors.lastname: Schaefer
authors.forename: Stefan
authors.initials: S
authors.lastname: Lorkowski
authors.forename: Siegmund
authors.initials: S
authors.lastname: Reissmann
mesh_terms: D000818:Animals; D002463:Cell Membrane Permeability / Q000235:genetics; D057846:Cell-Penetrating Peptides / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D004247:DNA / Q000235:genetics / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism / Q000633:toxicity; D007891:Leishmania / Q000235:genetics* / Q000378:metabolism / Q000472:pathogenicity; D014161:Transduction, Genetic; D001616:beta-Galactosidase / Q000378:metabolism
chemical_list: D057846:Cell-Penetrating Peptides; D004268:DNA-Binding Proteins; C475074:MPG peptide; D004247:DNA; D001616:beta-Galactosidase
keywords: CELL-PENETRATING PEPTIDES; CYTOTOXICITY; INTERNALIZATION OF PROTEINS; INTRACELLULAR CONCENTRATIONS; INTRACELLULAR DISTRIBUTION; LEISHMANIA TARENTOLAE, FORMATION OF NON-COVALENT COMPLEXES; UPTAKE EFFICIENCY