Mechanisms of peptide amphiphile internalization by SJSA-1 cells in vitro.

Self-assembly of peptide amphiphiles into nanostructures makes them attractive for a variety of applications in drug and peptide delivery. We here report on the interactions of micelles composed of a palmitoylated, pro-apoptotic peptide derived from p53 tumor suppressor protein with a human cancer cell line. Characterization of self-assembly in aqueous buffered solutions revealed formation of elongated rod-like micelles above a critical micelle concentration. Our results however demonstrate that monomers instead of micelles are internalized, a finding that correlates with the dynamic nature of the assemblies and the noncovalent interactions that hold them together. Internalization is shown to occur via adsorption-mediated, energy-dependent pathways, resulting in accumulation of the material in endocytic vesicles. We conclude that palmitoylation of peptides is an efficient way to increase peptide permeability inside SJSA-1 cells and that increased micelle stability would be required for intact micelle internalization.

[1]  José García de la Torre,et al.  Comparison of theories for the translational and rotational diffusion coefficients of rod‐like macromolecules. Application to short DNA fragments , 1984 .

[2]  R. M. Peitzsch,et al.  Binding of acylated peptides and fatty acids to phospholipid vesicles: pertinence to myristoylated proteins. , 1993, Biochemistry.

[3]  L. Smith,et al.  A nonexchangeable apolipoprotein E peptide that mediates binding to the low density lipoprotein receptor. , 1994, Journal of Biological Chemistry.

[4]  U. Ribel,et al.  Albumin binding of insulins acylated with fatty acids: characterization of the ligand-protein interaction and correlation between binding affinity and timing of the insulin effect in vivo. , 1995, The Biochemical journal.

[5]  G. Gerber,et al.  A direct role for serum albumin in the cellular uptake of long-chain fatty acids. , 1995, The Biochemical journal.

[6]  A. Levine,et al.  Structure of the MDM2 Oncoprotein Bound to the p53 Tumor Suppressor Transactivation Domain , 1996, Science.

[7]  S F Howard,et al.  Molecular characterization of the hdm2-p53 interaction. , 1997, Journal of molecular biology.

[8]  D. Reilly,et al.  A Sequence within the Cytoplasmic Tail of GpIIb Independently Activates Platelet Aggregation and Thromboxane Synthesis* , 1998, The Journal of Biological Chemistry.

[9]  B. Deurs,et al.  Extraction of cholesterol with methyl-beta-cyclodextrin perturbs formation of clathrin-coated endocytic vesicles. , 1999, Molecular biology of the cell.

[10]  K Kobylarz,et al.  Acute cholesterol depletion inhibits clathrin-coated pit budding. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[11]  C. Sergheraert,et al.  Direct evidence of cytoplasmic delivery of PKC‐α, ‐ϵ and ‐ζ pseudosubstrate lipopeptides: study of their implication in the induction of apoptosis , 1999 .

[12]  P. Furet,et al.  Discovery of potent antagonists of the interaction between human double minute 2 and tumor suppressor p53. , 2000, Journal of medicinal chemistry.

[13]  P. Dasgupta,et al.  Lipophilization of somatostatin analog RC‐160 with long chain fatty acid improves its antiproliferative and antiangiogenic activity in vitro , 2000, British journal of pharmacology.

[14]  A. Dautry‐Varsat,et al.  Endocytosis of an HIV‐derived lipopeptide into human dendritic cells followed by class I‐restricted CD8+ T lymphocyte activation , 2000, European journal of immunology.

[15]  L. Worley,et al.  Transducible peptide therapy for uveal melanoma and retinoblastoma. , 2002, Archives of ophthalmology.

[16]  J. Reig,et al.  Direct Visualization by Confocal Fluorescent Microscopy of the Permeation of Myristoylated Peptides Through the Cell Membrane , 2002, IUBMB life.

[17]  P. Chène.  Inhibiting the p53–MDM2 interaction: an important target for cancer therapy , 2003, Nature Reviews Cancer.

[18]  Eric Vives,et al.  Cell-penetrating Peptides , 2003, The Journal of Biological Chemistry.

[19]  Krista L. Niece,et al.  Selective Differentiation of Neural Progenitor Cells by High-Epitope Density Nanofibers , 2004, Science.

[20]  U. Ribel,et al.  The Mechanism of Protraction of Insulin Detemir, a Long-Acting, Acylated Analog of Human Insulin , 2004, Pharmaceutical Research.

[21]  R. Weissleder,et al.  Enhancing Membrane Permeability by Fatty Acylation of Oligoarginine Peptides , 2004, Chembiochem : a European journal of chemical biology.

[22]  N. Lockwood,et al.  Promotion of peptide antimicrobial activity by fatty acid conjugation. , 2004, Bioconjugate chemistry.

[23]  Ole G Mouritsen,et al.  Simulations of a membrane-anchored peptide: structure, dynamics, and influence on bilayer properties. , 2004, Biophysical journal.

[24]  Matthew Tirrell,et al.  Bottom-up design of biomimetic assemblies. , 2004, Advanced drug delivery reviews.

[25]  J. Tremblay,et al.  Endosome disruption enhances the functional nuclear delivery of Tat-fusion proteins. , 2004, Biochemical and biophysical research communications.

[26]  A. Caflisch,et al.  Membrane localization and flexibility of a lipidated ras peptide studied by molecular dynamics simulations. , 2004, Journal of the American Chemical Society.

[27]  B. Imperiali,et al.  The engineering of membrane-permeable peptides. , 2005, Analytical biochemistry.

[28]  M. Resh,et al.  Trafficking and signaling by fatty-acylated and prenylated proteins , 2006, Nature chemical biology.

[29]  Samuel I Stupp,et al.  Heparin binding nanostructures to promote growth of blood vessels. , 2006, Nano letters.

[30]  Badriprasad Ananthanarayanan,et al.  Increase of fluorescence anisotropy upon self-assembly in headgroup-labeled surfactants. , 2007, Macromolecular bioscience.

[31]  N. Allbritton,et al.  Myristoyl-based transport of peptides into living cells. , 2007, Biochemistry.

[32]  L. Vassilev,et al.  MDM2 inhibitors for cancer therapy. , 2007, Trends in molecular medicine.

[33]  P. Fischer.  Cellular uptake mechanisms and potential therapeutic utility of peptidic cell delivery vectors: Progress 2001–2006 , 2007, Medicinal research reviews.

[34]  John A. Robinson,et al.  Synthetic virus-like particles from self-assembling coiled-coil lipopeptides and their use in antigen display to the immune system. , 2007, Angewandte Chemie.

[35]  G. Fields,et al.  Targeted drug delivery utilizing protein-like molecular architecture. , 2007, Journal of the American Chemical Society.

[36]  Baoli Hu,et al.  Efficient p53 activation and apoptosis by simultaneous disruption of binding to MDM2 and MDMX. , 2007, Cancer research.

[37]  S. Gellman,et al.  Targeting protein-protein interactions: lessons from p53/MDM2. , 2007, Biopolymers.

[38]  S. Korsmeyer,et al.  Reactivation of the p53 Tumor Suppressor Pathway by a Stapled p53 Peptide , 2007 .

[39]  A. Mata,et al.  Self-Assembly of Large and Small Molecules into Hierarchically Ordered Sacs and Membranes , 2008, Science.

[40]  J. Schulz,et al.  Efficient Inhibition of the Alzheimer's Disease β-Secretase by Membrane Targeting , 2008, Science.

[41]  G. Roussel,et al.  Route of Uptake of Palmitoylated Encephalitogenic Peptides of Myelin Proteolipid Protein by Antigen-Presenting Cells: Importance of the Type of Bond between Lipid Chain and Peptide and Relevance to Autoimmunity1 , 2008, The Journal of Immunology.

[42]  G. D’Errico,et al.  Interaction of a peptide derived from glycoprotein gp36 of feline immunodeficiency virus and its lipoylated analogue with phospholipid membranes. , 2008, Biochemistry.

[43]  A. Kros,et al.  Scope and Applications of Amphiphilic Alkyl‐ and Lipopeptides , 2008 .

[44]  S. Futaki,et al.  Octaarginine- and Octalysine-modified Nanoparticles Have Different Modes of Endosomal Escape* , 2008, Journal of Biological Chemistry.

[45]  Felix Kratz,et al.  Albumin as a drug carrier: design of prodrugs, drug conjugates and nanoparticles. , 2008, Journal of controlled release : official journal of the Controlled Release Society.

[46]  Hans P Merkle,et al.  On the biomedical promise of cell penetrating peptides: limits versus prospects. , 2008, Journal of pharmaceutical sciences.

[47]  Y. Yoshioka,et al.  Organelle-targeted delivery of biological macromolecules using the protein transduction domain: potential applications for Peptide aptamer delivery into the nucleus. , 2008, Journal of molecular biology.

[48]  Erkki Ruoslahti,et al.  Targeting of albumin-embedded paclitaxel nanoparticles to tumors. , 2009, Nanomedicine : nanotechnology, biology, and medicine.