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Internalisation of cell-penetrating peptides into tobacco protoplasts
Stockholm University, Faculty of Science, Department of Neurochemistry and Neurotoxicology. Tallinn University of Technology, Estonia.
Stockholm University, Faculty of Science, Department of Neurochemistry and Neurotoxicology.
Stockholm University, Faculty of Science, Department of Neurochemistry and Neurotoxicology.
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2005 (English)In: Biochimica et Biophysica Acta, ISSN 0006-3002, Vol. 1669, no 2, 101-107 p.Article in journal (Refereed) Published
Abstract [en]

Cells are protected from the surrounding environment by plasma membrane which is impenetrable for most hydrophilic molecules. In the last 10 years cell-penetrating peptides (CPPs) have been discovered and developed. CPPs enter mammalian cells and carry cargo molecules over the plasma membrane with a molecular weight several times their own. Known transformation methods for plant cells have relatively low efficiency and require improvement. The possibility to use CPPs as potential delivery vectors for internalisation in plant cells has been studied in the present work. We analyse and compare the uptake of the fluorescein-labeled CPPs, transportan, TP10, penetratin and pVEC in Bowes human melanoma cells and Nicotiana tabacum cultivar (cv.) SR-1 protoplasts (plant cells without cell wall). We study the internalisation efficiency of CPPs with fluorescence microscopy, spectrofluorometry and fluorescence-activated cell sorter (FACS). All methods indicate, for the first time, that these CPPs can internalise into N. tabacum cv. SR-1 protoplasts. Transportan has the highest uptake efficacy among the studied peptides, both in mammalian cells and plant protoplast. The internalisation of CPPs by plant protoplasts may open up a new effective method for transfection in plants.

Place, publisher, year, edition, pages
2005. Vol. 1669, no 2, 101-107 p.
Keyword [en]
Cell-penetrating peptide, Plant protoplast, Membrane penetration, Transportan, TP10, Penetratin, pVEC
National Category
Biological Sciences Chemical Sciences
URN: urn:nbn:se:su:diva-24617DOI: 10.1016/j.bbamem.2005.01.006OAI: diva2:197938
Available from: 2008-01-10 Created: 2008-01-10 Last updated: 2015-04-21Bibliographically approved
In thesis
1. Cell-penetrating peptides in cargo delivery: In vitro studies on uptake and in vivo studies on tumor targeting
Open this publication in new window or tab >>Cell-penetrating peptides in cargo delivery: In vitro studies on uptake and in vivo studies on tumor targeting
2008 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

The capability of cell-penetrating peptides (CPPs) to transport cargos over different cellular membranes both in vitro and in vivo have drawn major attention in the past decade. Three main topics on the application of CPPs have been studied in this thesis.

First, several well-known CPPs, with fluorescein as a cargo, were shown to translocate into Nicotiana tabacum cultivar SR-1 protoplasts. By coupling different cargos to CPP it might be possible to effectively transport them inside the plant protoplasts. The translocation of CPPs into plant protoplasts might open up a new method for transformation of plant cells.

Next, the cell-penetrating ability of the novel peptide YTA2 was characterized and it was established that chemical coupling between YTA2 and the protein cargo is not needed for the transport of the cargo over the cellular membrane in vitro. The delivery of proteins into cells by mere coincubation with CPPs is an improvement, since the chemical coupling between the CPP and the cargo molecule, which adds time-consuming synthesis and purification steps, can be omitted.

Finally, by conjugating each of the two breast tumor homing peptides, the cyclic cCPGPEGAGC (PEGA) or the linear CREKA peptide, to the CPP pVEC, two cell-penetrating peptides with homing properties were obtained. Both, PEGA-pVEC and CREKA-pVEC were taken up by different breast cancer cells in vitro. Moreover, the homing capacity of the PEGA-pVEC and CREKA-pVEC was conserved in vivo, where the conjugates mainly accumulated in blood vessels in breast tumor tissue and were subsequently translocated into cells. Conjugating the anti-cancer drug chlorambucil to PEGA-pVEC or CREKA-pVEC markedly improved its efficiency. Furthermore, systemic treatment of tumor-bearing mice with chlorambucil-CREKA-pVEC significantly reduced tumor growth compared to control groups. These tumor-homing CPPs might improve both diagnosis and treatment of breast cancer tumors, by conjugation to therapeutic agents.

Place, publisher, year, edition, pages
Stockholm: Institutionen för neurokemi, 2008. 67 p.
National Category
Research subject
Neurochemistry with Molecular Neurobiology
urn:nbn:se:su:diva-7263 (URN)978-91-7155-554-0 (ISBN)
Public defence
2008-02-01, Magnélisalen, Kemiska övningslaboratoriet, Svante Arrhenius väg 12 A, Stockholm, 10:00
Available from: 2008-01-10 Created: 2008-01-10 Last updated: 2015-03-23Bibliographically approved

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Langel, Ülo
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