Synthesis by chemoselective ligation and biological evaluation of novel cell-permeable PKC-zeta pseudosubstrate lipopeptides.
The ability of lipopeptides to passively cross the cell membrane opens new opportunities for the intracellular delivery of bioactive peptides. However, the production of large series of cell-permeable lipopeptides is not trivial due to their generally low solubility. We have evaluated the possibility of associating the fatty acid to the functional cargo using generally applicable ligation chemistries. To this end, we have designed an amphiphilic shuttle in which arginine residues served to solubilize the lipid part in aqueous media, during both the assembly of the lipopeptide and the cellular assays. Our model peptide, the pseudosubstrate sequence of protein kinase C-zeta (PKC-zeta), was associated to the pentapeptide Gly-Arg-Gly-Arg-Lys(Pam)-NH2 through thiazolidine, thioether, disulfide, or hydrazone linkages. The cytoplasm import of the resulting constructs was monitored through the quantification of the apoptosis specifically induced by PKC-zeta inhibition. Our observations suggested the interest of this noninvasive cellular import method to modulate the activity of an intracytoplasmic pharmacological target and showed the influence of a non-amide link created between the functional peptide and the lipidic vector: optimal results, in terms of both specific activity and low basal cytotoxicity, were obtained with the thiazolidine ligation product.
Bonnet
D
D
Institut of Biology and Pasteur Institute of Lille, University of Lille II, UMR 8525 CNRS, France.
Thiam
K
K
Loing
E
E
Melnyk
O
O
Gras-Masse
H
H
D017209:Apoptosis; D002463:Cell Membrane Permeability; D004220:Disulfides / Q000737:chemistry; D006801:Humans; D006835:Hydrazones / Q000737:chemistry; D019169:Jurkat Cells; D009842:Oligopeptides / Q000138:chemical synthesis* / Q000737:chemistry / Q000494:pharmacology; D019308:Palmitic Acid / Q000737:chemistry*; D011493:Protein Kinase C / Q000737:chemistry*; D013440:Sulfides / Q000737:chemistry; D013844:Thiazoles / Q000737:chemistry; D000099221:Protein Kinase C zeta
D004220:Disulfides; D006835:Hydrazones; D009842:Oligopeptides; D019308:Palmitic Acid; D011493:Protein Kinase C; D013440:Sulfides; D013844:Thiazoles; D000099221:Protein Kinase C zeta