Intracellular Regulation of Macromolecules Using pH-Sensitive Liposomes and Nuclear Localization Signal: Qualitative and Quantitative Evaluation of Intracellular Trafficking
| Autor: | Mineo Niwa, Hiroshi Kiwada, S. Futaki, Hideyoshi Harashima, Makiko Azumano, Rieko Tachibana, Masayuki Shono |
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| Rok vydání: | 1998 |
| Předmět: |
Male
Endosome Endocytic cycle Biophysics Endosomes Biology Endocytosis Biochemistry law.invention Confocal microscopy law Animals NLS Rats Wistar Molecular Biology Cell Nucleus Drug Carriers Liposome Phosphatidylethanolamines Serum Albumin Bovine Cell Biology Hydrogen-Ion Concentration Rats Cell biology Liposomes Macrophages Peritoneal Cattle Cholesterol Esters Nuclear transport Fluorescein-5-isothiocyanate Nuclear localization sequence |
| Zdroj: | Biochemical and Biophysical Research Communications. 251:538-544 |
| ISSN: | 0006-291X |
| Popis: | The objective of this study is to present a rational strategy to target macromolecules to the nucleus via the endocytic pathway. The two major barriers in this route to the nucleus are known as endosomal escape and nuclear transport. pH-sensitive liposomes were used in order to achieve endosomal escape under the conditions of low pH in endosomes. Bovine serum albumin (alb) served as a model compound to be delivered to nucleus and was encapsulated into the pH-sensitive liposomes. The liposomes are composed of dioleoyl phosphatidyl ethanolamine: cholesterylhemisuccinate. They were taken up by rat peritoneal macrophages via endocytosis and subsequently underwent degradation, principally by lysosomal enzymes. By using pH-sensitive liposomes, intracellular degradation was reduced by a significant extent, as expected, via endosomal escape. Cytosolic delivery of FITC-labelled alb was also detected by confocal microscopy. Selective targeting to the nucleus was performed by adding the nuclear localization signal (NLS) of the SV-40 large T antigen to the FITC-alb, which were then encapsulated into the pH-sensitive liposomes. Confocal microscopy revealed that FITC-alb, in the presence of NLS was successfully delivered into nucleus, while no transport was observed in the absence of NLS. These results provide a useful strategy for the nuclear targeting of macromolecules using pH-sensitive liposomes in conjunction with NLS. |
| Databáze: | OpenAIRE |
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