Document Detail

Efficient delivery of cell impermeable phosphopeptides by a cyclic peptide amphiphile containing tryptophan and arginine.
MedLine Citation:
PMID:  23537165     Owner:  NLM     Status:  MEDLINE    
Phosphopeptides are valuable reagent probes for studying protein-protein and protein-ligand interactions. The cellular delivery of phosphopeptides is challenging because of the presence of the negatively charged phosphate group. The cellular uptake of a number of fluorescent-labeled phosphopeptides, including F'-GpYLPQTV, F'-NEpYTARQ, F'-AEEEIYGEFEAKKKK, F'-PEpYLGLD, F'-pYVNVQN-NH2, and F'-GpYEEI (F' = fluorescein), was evaluated in the presence or absence of a [WR]4, a cyclic peptide containing alternative arginine (R) and tryptophan (W) residues, in human leukemia cells (CCRF-CEM) after 2 h incubation using flow cytometry. [WR]4 improved significantly the cellular uptake of all phosphopeptides. PEpYLGLD is a sequence that mimics the pTyr1246 of ErbB2 that is responsible for binding to the Chk SH2 domain. The cellular uptake of F'-PEpYLGLD was enhanced dramatically by 27-fold in the presence of [WR]4 and was found to be time-dependent. Confocal microscopy of a mixture of F'-PEpYLGLD and [WR]4 in live cells exhibited intracellular localization and significantly higher cellular uptake compared to that of F'-PEpYLGLD alone. Transmission electron microscopy (TEM) and isothermal calorimetry (ITC) were used to study the interaction of PEpYLGLD and [WR]4. TEM results showed that the mixture of PEpYLGLD and [WR]4 formed noncircular nanosized structures with width and height of 125 and 60 nm, respectively. ITC binding studies confirmed the interaction between [WR]4 and PEpYLGLD. The binding isotherm curves, derived from sequential binding models, showed an exothermic interaction driven by entropy. These studies suggest that amphiphilic peptide [WR]4 can be used as a cellular delivery tool of cell-impermeable negatively charged phosphopeptides.
Amir Nasrolahi Shirazi; Rakesh Kumar Tiwari; Donghoon Oh; Antara Banerjee; Arpita Yadav; Keykavous Parang
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, U.S. Gov't, Non-P.H.S.     Date:  2013-04-15
Journal Detail:
Title:  Molecular pharmaceutics     Volume:  10     ISSN:  1543-8392     ISO Abbreviation:  Mol. Pharm.     Publication Date:  2013 May 
Date Detail:
Created Date:  2013-05-06     Completed Date:  2013-12-16     Revised Date:  2014-05-08    
Medline Journal Info:
Nlm Unique ID:  101197791     Medline TA:  Mol Pharm     Country:  United States    
Other Details:
Languages:  eng     Pagination:  2008-20     Citation Subset:  IM    
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MeSH Terms
Amino Acid Sequence
Arginine / chemistry
Biological Transport, Active
Cell Line, Tumor
Cell Membrane Permeability
Drug Carriers / chemistry
Drug Delivery Systems
Fluorescent Dyes / chemistry
Molecular Structure
Peptides, Cyclic / chemistry
Phosphopeptides / administration & dosage*,  chemistry*,  pharmacokinetics
Protein Interaction Domains and Motifs
Surface-Active Agents / chemistry
Tryptophan / chemistry
Grant Support
8 P20 GM103430-12/GM/NIGMS NIH HHS; P20 RR016457/RR/NCRR NIH HHS
Reg. No./Substance:
0/Drug Carriers; 0/Fluorescent Dyes; 0/Peptides, Cyclic; 0/Phosphopeptides; 0/Surface-Active Agents; 8DUH1N11BX/Tryptophan; 94ZLA3W45F/Arginine

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

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