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Relationships between Cargo, Cell Penetrating Peptides and Cell Type for Uptake of Non-Covalent Complexes into Live Cells
Biological and Pharmaceutical Faculty, Friedrich Schiller University, Dornburger Str. 25, 07743 Jena, Germany
Jena Bioscience GmbH, Loebstedter Str. 80, 07749 Jena, Germany
Leibniz Institute for Age Research - Fritz Lipmann Institute, Beutenbergstr. 11, 07745 Jena, Germany
* Author to whom correspondence should be addressed.
Received: 1 November 2012; in revised form: 29 January 2013 / Accepted: 30 January 2013 / Published: 6 February 2013
Abstract: Modulating signaling pathways for research and therapy requires either suppression or expression of selected genes or internalization of proteins such as enzymes, antibodies, nucleotide binding proteins or substrates including nucleoside phosphates and enzyme inhibitors. Peptides, proteins and nucleotides are transported by fusing or conjugating them to cell penetrating peptides or by formation of non-covalent complexes. The latter is often preferred because of easy handling, uptake efficiency and auto-release of cargo into the live cell. In our studies complexes are formed with labeled or readily detectable cargoes for qualitative and quantitative estimation of their internalization. Properties and behavior of adhesion and suspension vertebrate cells as well as the protozoa Leishmania tarentolae are investigated with respect to proteolytic activity, uptake efficiency, intracellular localization and cytotoxicity. Our results show that peptide stability to membrane-bound, secreted or intracellular proteases varies between different CPPs and that the suitability of individual CPPs for a particular cargo in complex formation by non-covalent interactions requires detailed studies. Cells vary in their sensitivity to increasing concentrations of CPPs. Thus, most cells can be efficiently transduced with peptides, proteins and nucleotides with intracellular concentrations in the low micromole range. For each cargo, cell type and CPP the optimal conditions must be determined separately.
Keywords: cell penetrating peptides; CPPs; formation of non-covalent complexes; MPG peptides; internalized amount of cargoes; relationships between CPP, cargo and cell type
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Cite This Article
MDPI and ACS Style
Keller, A.-A.; Mussbach, F.; Breitling, R.; Hemmerich, P.; Schaefer, B.; Lorkowski, S.; Reissmann, S. Relationships between Cargo, Cell Penetrating Peptides and Cell Type for Uptake of Non-Covalent Complexes into Live Cells. Pharmaceuticals 2013, 6, 184-203.
Keller A-A, Mussbach F, Breitling R, Hemmerich P, Schaefer B, Lorkowski S, Reissmann S. Relationships between Cargo, Cell Penetrating Peptides and Cell Type for Uptake of Non-Covalent Complexes into Live Cells. Pharmaceuticals. 2013; 6(2):184-203.
Keller, Andrea-Anneliese; Mussbach, Franziska; Breitling, Reinhard; Hemmerich, Peter; Schaefer, Buerk; Lorkowski, Stefan; Reissmann, Siegmund. 2013. "Relationships between Cargo, Cell Penetrating Peptides and Cell Type for Uptake of Non-Covalent Complexes into Live Cells." Pharmaceuticals 6, no. 2: 184-203.