Antibody caging of a nuclear-targeting signal.
We have developed a technique for reversibly masking a peptide-targeting signal. A fluoresceinated derivative of the simian virus 40 large tumor antigen nuclear-targeting signal was synthesized and cross-linked to bovine serum albumin. The conjugated protein was efficiently transported into rat liver nuclei unless the peptide-targeting signal was sterically hindered by binding of an anti-fluorescein antibody. Addition of free 5-aminofluorescein competed for antibody binding and rapidly restored nuclear accumulation of the derivatized bovine serum albumin. General use of hapten derivatization and anti-hapten antibodies for caging portions of macromolecular surfaces can be extended to a variety of proteins, including antibodies themselves.
Halleck
M S
MS
Department of Biochemistry, University of Utah School of Medicine, Salt Lake City 84132.
Rechsteiner
M
M
D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies; D000911:Antibodies, Monoclonal; D000952:Antigens, Polyomavirus Transforming; D002467:Cell Nucleus / Q000378:metabolism*; D005260:Female; D016650:Fluorescein-5-isothiocyanate; D005452:Fluoresceins; D007700:Kinetics; D008099:Liver / Q000378:metabolism; D008969:Molecular Sequence Data; D009865:Oocytes / Q000378:metabolism; D010455:Peptides / Q000276:immunology; D051381:Rats; D015398:Signal Transduction; D013539:Simian virus 40 / Q000276:immunology; D013861:Thiocyanates; D014357:Trypsin; D014982:Xenopus laevis
D000906:Antibodies; D000911:Antibodies, Monoclonal; D000952:Antigens, Polyomavirus Transforming; D005452:Fluoresceins; D010455:Peptides; D013861:Thiocyanates; D014357:Trypsin; D016650:Fluorescein-5-isothiocyanate