Internalisation of cell-penetrating peptides into tobacco protoplasts.
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.
Mäe
Maarja
M
Department of Neurochemistry and Neurotoxicology, Stockholm University, Svante Arrhenius väg 21A, SE-106 91 Stockholm, Sweden. maarja@neurochem.su.se
Myrberg
Helena
H
Jiang
Yang
Y
Paves
Heiti
H
Valkna
Andres
A
Langel
Ulo
U
D002352:Carrier Proteins / Q000378:metabolism*; D057846:Cell-Penetrating Peptides; D005434:Flow Cytometry; D019793:Fluorescein; D019004:Galanin; D006801:Humans; D010455:Peptides / Q000378:metabolism*; D021381:Protein Transport; D011523:Protoplasts / Q000378:metabolism*; D011951:Receptors, Complement; D011993:Recombinant Fusion Proteins / Q000378:metabolism*; D013050:Spectrometry, Fluorescence; D014026:Nicotiana / Q000378:metabolism*; D014407:Tumor Cells, Cultured; D014862:Wasp Venoms
D002352:Carrier Proteins; D057846:Cell-Penetrating Peptides; D010455:Peptides; D011951:Receptors, Complement; D011993:Recombinant Fusion Proteins; D014862:Wasp Venoms; C437783:pVEC peptide; C110068:transportan; C082885:soluble complement inhibitor 1; D019004:Galanin; C414409:penetratin; D019793:Fluorescein