Bovine serum albumin is a major oligonucleotide-binding protein found on the surface of cultured cells.
To better understand the uptake of oligonucleotides into cells, we have studied the labeling of cell surface proteins by an oligonucleotide conjugated to a radiolabeled photoactivatable crosslinker (Denny-Jaffe reagent). When HL60 cells are treated with the conjugate for 2 hours in a medium containing bovine serum albumin (BSA), almost all of the cell-associated label is found in one protein, which we identify as BSA. Cells grown and treated in a serum-free medium do not show this protein, whereas it is plainly seen in cells that are grown in serum-containing medium but then treated in serum-free medium. Overall association of the oligonucleotide with cells is much higher in serum-free medium than in BSA-containing medium, but the oligonucleotide is mostly not protein-associated in the absence of BSA. We conclude that (1) BSA from the medium serves to block overall association of oligonucleotide with cells, and (2) BSA is the main cell surface protein binding oligonucleotides. We discuss the possible role of albumin in endocytic uptake of oligonucleotides in the cell and in the biodistribution of oligonucleotides in vivo.
Geselowitz
D A
DA
Department of Nuclear Medicine, National Institutes of Health, Bethesda, Maryland 20892-1180, USA.
Neckers
L M
LM
D000818:Animals; D001483:Base Sequence; D002352:Carrier Proteins / Q000032:analysis / Q000378:metabolism; D002417:Cattle; D002462:Cell Membrane / Q000378:metabolism; D003470:Culture Media; D016895:Culture Media, Serum-Free; D004797:Enzyme-Linked Immunosorbent Assay; D018922:HL-60 Cells; D006801:Humans; D008969:Molecular Sequence Data; D016376:Oligonucleotides, Antisense / Q000138:chemical synthesis / Q000378:metabolism* / Q000493:pharmacokinetics; D012710:Serum Albumin, Bovine / Q000032:analysis / Q000378:metabolism*
D002352:Carrier Proteins; D003470:Culture Media; D016895:Culture Media, Serum-Free; D016376:Oligonucleotides, Antisense; D012710:Serum Albumin, Bovine