title: Guanidinylated neomycin delivers large, bioactive cargo into cells through a heparan sulfate-dependent pathway.
abstract: Facilitating the uptake of molecules into living cells is of substantial interest for basic research and drug delivery applications. Arginine-rich peptides have been shown to facilitate uptake of high molecular mass cargos into cells, but the mechanism of uptake is complex and may involve multiple receptors. In this report, we show that a derivative of the aminoglycoside antibiotic neomycin, in which all of the ammonium groups have been converted into guanidinium groups, can carry large (>300 kDa) bioactive molecules across cell membranes. Delivery occurs at nanomolar transporter concentrations and under these conditions depends entirely on cell surface heparan sulfate proteoglycans. Conjugation of guanidinoneomycin to the plant toxin saporin, a ribosome-inactivating agent, results in proteoglycan-dependent cell toxicity. In contrast, an arginine-rich peptide shows both heparan sulfate-dependent and -independent cellular uptake. The high selectivity of guanidinoneomycin for heparan sulfate suggests the possibility of exploiting differences in proteoglycan compositions to target delivery to different cell types.
authors.affiliation: Department of Chemistry and Biochemistry and Cellular, University of California, San Diego, La Jolla, California 92093, USA.
authors.forename: Lev
authors.initials: L
authors.lastname: Elson-Schwab
authors.forename: Omai B
authors.initials: OB
authors.lastname: Garner
authors.forename: Manuela
authors.initials: M
authors.lastname: Schuksz
authors.forename: Brett E
authors.initials: BE
authors.lastname: Crawford
authors.forename: Jeffrey D
authors.initials: JD
authors.lastname: Esko
authors.forename: Yitzhak
authors.initials: Y
authors.lastname: Tor
mesh_terms: D000818:Animals; D001692:Biological Transport / Q000187:drug effects / Q000502:physiology; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D004305:Dose-Response Relationship, Drug; D016503:Drug Delivery Systems; D019791:Guanidine / Q000031:analogs & derivatives / Q000138:chemical synthesis / Q000493:pharmacokinetics; D019812:Heparan Sulfate Proteoglycans / Q000378:metabolism*; D008562:Membrane Glycoproteins / Q000378:metabolism*; D009699:N-Glycosyl Hydrolases / Q000493:pharmacokinetics; D009355:Neomycin / Q000031:analogs & derivatives / Q000138:chemical synthesis / Q000493:pharmacokinetics*; D010455:Peptides / Q000138:chemical synthesis / Q000493:pharmacokinetics; D010940:Plant Proteins / Q000493:pharmacokinetics; D011485:Protein Binding / Q000187:drug effects; D011500:Protein Synthesis Inhibitors / Q000138:chemical synthesis / Q000493:pharmacokinetics*; D054789:Ribosome Inactivating Proteins, Type 1; D000076985:Saporins
chemical_list: D019812:Heparan Sulfate Proteoglycans; D008562:Membrane Glycoproteins; D010455:Peptides; D010940:Plant Proteins; D011500:Protein Synthesis Inhibitors; D054789:Ribosome Inactivating Proteins, Type 1; D009699:N-Glycosyl Hydrolases; D000076985:Saporins; D009355:Neomycin; D019791:Guanidine