Rational development of caged-biotin protein-labeling agents and some applications in live cells.
Biotin-(strept)avidin complex is widely used in biotechnology because of its extremely high binding constant, but there is no report describing spatiotemporally controlled formation of the complex in live cells. Here, based on X-ray crystal structure analysis and calorimetric data, we designed and synthesized photoreleasable biotins, which show greatly reduced affinity for (strept)avidin, but recover native affinity after UV irradiation. For application at the cell surface, we introduced an amine-reactive moiety into these "caged" biotin molecules. Specific fluorescence imaging of live cells that had been labeled with these agents and then UV-irradiated, was accomplished by addition of streptavidin conjugated with a fluorophore. We also demonstrated the applicability of these compounds for UV-irradiated-cell-specific drug delivery by using caged-biotin-labeled cells, a prodrug, and streptavidin conjugated with a prodrug-activating enzyme.
Graduate School of Pharmaceutical Sciences, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan.
Takuya
T
Terai
Eri
E
Maki
Shigeru
S
Sugiyama
Yoshinori
Y
Takahashi
Hiroyoshi
H
Matsumura
Yusuke
Y
Mori
Tetsuo
T
Nagano
D001360:Avidin / Q000737:chemistry / Q000378:metabolism*; D001710:Biotin / Q000737:chemistry*; D006690:Bisbenzimidazole / Q000737:chemistry / Q000378:metabolism; D016923:Cell Death / Q000187:drug effects / Q000528:radiation effects; D002467:Cell Nucleus / Q000737:chemistry / Q000378:metabolism; D018360:Crystallography, X-Ray; D004317:Doxorubicin / Q000494:pharmacology; D016503:Drug Delivery Systems; D005456:Fluorescent Dyes / Q000737:chemistry; D006367:HeLa Cells / Q000187:drug effects / Q000528:radiation effects; D006801:Humans; D008239:Lysine / Q000737:chemistry; D008565:Membrane Proteins / Q000737:chemistry; D018613:Microscopy, Confocal; D015394:Molecular Structure; D010777:Photochemistry / Q000379:methods*; D011355:Prodrugs / Q000494:pharmacology; D015202:Protein Engineering / Q000379:methods*; D011994:Recombinant Proteins / Q000737:chemistry* / Q000378:metabolism; D013050:Spectrometry, Fluorescence; D019809:Streptavidin / Q000737:chemistry / Q000378:metabolism; D014466:Ultraviolet Rays; D014508:Urea / Q000737:chemistry
D005456:Fluorescent Dyes; D008565:Membrane Proteins; D011355:Prodrugs; D011994:Recombinant Proteins; D001360:Avidin; D001710:Biotin; D004317:Doxorubicin; D014508:Urea; D019809:Streptavidin; D008239:Lysine; D006690:Bisbenzimidazole