Uptake by macrophages of a biotinylated oligo-alpha-deoxythymidylate by using mannosylated streptavidin.
Streptavidin substituted with mannose residues increased by 20-fold the intracellular concentration of a biotinylated dodecakis(alpha-deoxythymidylate) in macrophages by comparison with the uptake of free oligodeoxynucleotide. Streptavidin, the bacterial homologue of the very basic avidin, which does not contain any carbohydrate moieties and is a neutral protein, was substituted with 12 mannose residues in order to be recognized and internalized by mannose-specific lectins on the surface of macrophages. A 3'-biotinylated and 5'-fluoresceinylated dodecakis (alpha-deoxythymidylate) was synthesized and bound onto mannosylated streptavidin. The conjugate was isolated, and by using flow cytometry, it was shown that the uptake of fluoresceinylated oligodeoxynucleotides bound to mannosylated streptavidin by macrophages is 20-fold higher than that of free oligodeoxynucleotides and that the uptake was competively inhibited by mannosylated serum albumin. Glycosylated streptavidin conjugates recognizing specific membrane lectins on different cells provide the possibility to target biotinylated antisense oligodeoxynucleotides and to increase the biological effect of these chemotherapeutic agents.
Bonfils
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Laboratoire de Biochimie des Glycoconjugués et Lectines endogènes, CNRS, Orléans, France.
Mendes
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Roche
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AC
Monsigny
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Midoux
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P
D000818:Animals; D001426:Bacterial Proteins / Q000138:chemical synthesis / Q000737:chemistry*; D001710:Biotin / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000378:metabolism; D002460:Cell Line; D004591:Electrophoresis, Polyacrylamide Gel; D004705:Endocytosis; D005434:Flow Cytometry; D008264:Macrophages / Q000378:metabolism*; D008362:Mannosides / Q000138:chemical synthesis / Q000737:chemistry*; D051379:Mice; D009838:Oligodeoxyribonucleotides / Q000138:chemical synthesis / Q000378:metabolism*; D019809:Streptavidin; D013938:Thymidine Monophosphate / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000378:metabolism
D001426:Bacterial Proteins; D008362:Mannosides; D009838:Oligodeoxyribonucleotides; C027903:oligo (dT); C077294:3'-biotinyl-5'-fluorosceinyldodecakis(alpha-deoxythymidylate); D013938:Thymidine Monophosphate; D001710:Biotin; D019809:Streptavidin