Crotamine mediates gene delivery into cells through the binding to heparan sulfate proteoglycans.
Recently we have shown that crotamine, a toxin from the South American rattlesnake Crotalus durissus terrificus venom, belongs to the family of cell-penetrating peptides. Moreover, crotamine was demonstrated to be a marker of centrioles, of cell cycle, and of actively proliferating cells. Herein we show that this toxin at non-toxic concentrations is also capable of binding electrostatically to plasmid DNA forming DNA-peptide complexes whose stabilities overcome the need for chemical conjugation for carrying nucleic acids into cells. Interestingly, crotamine demonstrates cell specificity and targeted delivery of plasmid DNA into actively proliferating cells both in vitro and in vivo, which distinguishes crotamine from other known natural cell-penetrating peptides. The mechanism of crotamine penetration and cargo delivery into cells was also investigated, showing the involvement of heparan sulfate proteoglycans in the uptake phase, which is followed by endocytosis and peptide accumulation within the acidic endosomal vesicles. Finally, the permeabilization of endosomal membranes induced by crotamine results in the leakage of the vesicles contents to the cell cytosol.
Nascimento
Fábio Dupart
FD
Departamento de Bioquímica, Universidade Federal de São Paulo-Escola Paulista de Medicina, Rua 3 de Maio, 100, Ed. INFAR, CEP 04044-020, São Paulo, Brazil.
Hayashi
Mirian A F
MA
Kerkis
Alexandre
A
Oliveira
Vitor
V
Oliveira
Eduardo B
EB
Rádis-Baptista
Gandhi
G
Nader
Helena Bonciani
HB
Yamane
Tetsuo
T
Tersariol
Ivarne Luis dos Santos
IL
Kerkis
Irina
I
D000818:Animals; D016466:CHO Cells; D002462:Cell Membrane / Q000378:metabolism; D049109:Cell Proliferation; D006224:Cricetinae; D003412:Cricetulus; D003435:Crotalid Venoms / Q000494:pharmacology*; D053595:Embryonic Stem Cells / Q000166:cytology; D004705:Endocytosis; D011992:Endosomes / Q000378:metabolism; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D019812:Heparan Sulfate Proteoglycans / Q000737:chemistry* / Q000378:metabolism; D051379:Mice; D011509:Proteoglycans / Q000378:metabolism
D003435:Crotalid Venoms; D019812:Heparan Sulfate Proteoglycans; D011509:Proteoglycans; C010563:crotamine