Stimulated endocytosis in penetratin uptake: effect of arginine and lysine.
Cell-penetrating peptides can deliver macromolecular cargo into cells and show promise as vectors for intracellular drug delivery. Internalization occurs predominantly via endocytosis, but the exact uptake mechanisms are not fully understood. We show quantitatively how penetratin, a 16-residue cationic peptide, stimulates fluid-phase endocytosis and triggers its own uptake into Chinese hamster ovarian cells, using a 70kDa dextran to indicate macropinocytosis. The total cellular endocytotic rate is significantly less affected and we therefore propose up-regulation of macropinocytosis to occur at the expense of other types of endocytosis. By comparing penetratin to its analogs PenArg and PenLys, enriched in arginines and lysines, respectively, we show how these side-chains contribute to uptake efficiency. The degree of peptide and dextran uptake follows similar patterns regarding peptide concentration and arginine/lysine content (PenArg>penetratin>PenLys), indicating that a high content of arginines is beneficial but not necessary for stimulating endocytosis.
Amand
Helene L
HL
Department of Chemical and Biological Engineering/Physical Chemistry, Chalmers University of Technology, Kemivägen 10, S-412 96 Gothenburg, Sweden.
Fant
Kristina
K
Nordén
Bengt
B
Esbjörner
Elin K
EK
D000595:Amino Acid Sequence; D000818:Animals; D001120:Arginine / Q000737:chemistry*; D016466:CHO Cells; D002352:Carrier Proteins / Q000032:analysis / Q000737:chemistry* / Q000378:metabolism*; D057846:Cell-Penetrating Peptides; D006224:Cricetinae; D003412:Cricetulus; D003911:Dextrans / Q000378:metabolism; D004705:Endocytosis / Q000187:drug effects; D008239:Lysine / Q000737:chemistry*; D018613:Microscopy, Confocal; D008969:Molecular Sequence Data; D021381:Protein Transport
D002352:Carrier Proteins; D057846:Cell-Penetrating Peptides; D003911:Dextrans; D001120:Arginine; C414409:penetratin; D008239:Lysine