Cell Penetrating Peptide-Mediated Caveolae-Dependent Activation of Lung Endothelial Nitric Oxide Synthase
| Autor: | Tarun E. Hutchinson, Jawaharlal M. Patel, Hanbo Hu |
|---|---|
| Rok vydání: | 2016 |
| Předmět: |
0106 biological sciences
Nitric Oxide Synthase Type III Endothelium Swine media_common.quotation_subject Guanosine Cell-Penetrating Peptides 02 engineering and technology Caveolae Nitric Oxide 01 natural sciences Biochemistry Nitric oxide Phosphatidylinositol 3-Kinases chemistry.chemical_compound Structural Biology Enos 010608 biotechnology medicine Animals Humans Phosphorylation Internalization Lung media_common biology Kinase Endothelial Cells General Medicine 021001 nanoscience & nanotechnology biology.organism_classification Cell biology medicine.anatomical_structure chemistry Endothelium Vascular 0210 nano-technology Intracellular Signal Transduction |
| Zdroj: | Protein & Peptide Letters. 23:169-175 |
| ISSN: | 0929-8665 |
| Popis: | Cell penetrating peptides can be used as therapeutic agents via modulation of selective cell functions. Nitric oxide (NO) generated by vascular endothelial NO synthase (eNOS) plays a critical role in the NO/ cyclic guanosine 5'-monophosphate (cGMP)-mediated pulmonary vascular function. Here we examined whether internalization of a fifteen amino acid (KRFNSISCSSWRRKR) synthetic peptide (P3) enhances the catalytic activity of eNOS via caveolae/eNOS dissociation leading to NO release and increased cGMP production in pulmonary artery endothelial cells (EC). ECs were treated with varying concentrations of P3 and used to monitor internalization, isolation of caveolae-enriched fraction, the catalytic activity of eNOS, NO/cGMP production, and intracellular Ca(2+) release. Confocal images show timedependent internalization of P3 in EC. Treatment of EC with P3, but not scrambled P3, increased the catalytic activity of eNOS in a dose-dependent manner without change in eNOS expression or phosphorylation. Treatment of EC with P3 stimulated intracellular Ca(2+) release, increased the catalytic activity of phospatidylinsositide 3 kinase (PI3K) and resulted in eNOS/caveolae-1 (Cav-1) dissociation leading to translocation of eNOS to intracellular compartment in EC. P3- mediated activation of eNOS was abolished by intracellular Ca(2+) chelator 1,2-bis(2-aminophenooxy)ethane-N,N,N',N'- tertraacetic acid-AM (BAPTA-AM), PI3K inhibition, or by siRNA-mediated Cav-1 suppression. These results demonstrate that exogenous peptide consisting of cationic amino acids can internalize and enhance the catalytic activity of eNOS via modulation of caveolar signaling and intracellular Ca(2+) release in EC. |
| Databáze: | OpenAIRE |
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