Specific redistribution of cell-penetrating peptides from endosomes to the cytoplasm and nucleus upon laser illumination.
Cell-penetrating peptides (CPPs), once postulated to cross cell membranes in a non-endocytic, non-energy-dependent process, have since been found to accumulate in vesicles in live mammalian cells. In this study, we show that it is possible to use laser light from a confocal microscope to cause labeled peptide-conjugated CPPs to redistribute from vesicles into the cytoplasm and nucleus of cells. Following redistribution, the cells are found to be biologically responsive, and they retain morphology for several hours. It was possible to initiate redistribution of both fluorescein- and Alexa633-labeled peptides by selective irradiation of one of the fluorophores. These peptides could potentially be used as tracers to selectively deliver cargo biomolecules into cells by laser illumination using a standard fluorescence confocal microscope.
Maiolo
James R
JR
Department of Automated Biotechnology, Merck & Co. Inc., 502 Louise Lane, North Wales, PA 19454, USA.
Ottinger
Elizabeth A
EA
Ferrer
Marc
M
D045744:Cell Line, Tumor; D002463:Cell Membrane Permeability; D002467:Cell Nucleus / Q000378:metabolism*; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000737:chemistry / Q000378:metabolism; D003593:Cytoplasm / Q000378:metabolism; D011992:Endosomes / Q000378:metabolism*; D019793:Fluorescein / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D018613:Microscopy, Confocal; D012516:Osteosarcoma / Q000378:metabolism; D010446:Peptide Fragments / Q000737:chemistry / Q000378:metabolism*; D018343:Receptors, Adrenergic, beta-2 / Q000096:biosynthesis / Q000378:metabolism
C505032:CFTR protein, human; D010446:Peptide Fragments; D018343:Receptors, Adrenergic, beta-2; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D019793:Fluorescein