Synthesis and application of cleavable photoactivable heterobifunctional reagents.
Three different cleavable photoactivable heterobifunctional reagents have been synthesized and characterized by proton NMR, 13C NMR, mass spectroscopy, infrared spectroscopy, and thin-layer chromatography. They are the N-hydroxysuccinimide ester of 3-[(4-azidophenyl)dithio]-propionic acid, methyl 3-[(4-azidophenyl)dithio]propionimidate, and N-[(4-azidophenyl)thio]phthalimide. Concanavalin A was coupled with one of the reagents, the N-hydroxysuccinimide ester of 3-[(4-azidophenyl)dithio]propionic acid, and radioiodinated. The [125I]lectin derivative and [125I]lectin showed similar specific binding to the receptors on the human erythrocyte membrane. Upon photolysis, subunits of the [125I]lectin derivative were cross-linked to produce dimer, trimer, and tetramer. These cross-linked complexes were readily cleaved by reducing the disulfide bond of the reagent.
D001386:Azides; D003208:Concanavalin A; D003432:Cross-Linking Reagents / Q000138:chemical synthesis*; D004910:Erythrocyte Membrane / Q000276:immunology; D006801:Humans; D007096:Imidoesters / Q000138:chemical synthesis* / Q000494:pharmacology; D009682:Magnetic Resonance Spectroscopy; D013058:Mass Spectrometry; D010797:Phthalimides / Q000138:chemical synthesis* / Q000494:pharmacology; D011952:Receptors, Concanavalin A / Q000187:drug effects / Q000378:metabolism; D013056:Spectrophotometry, Ultraviolet; D013388:Succinimides / Q000138:chemical synthesis* / Q000494:pharmacology
D001386:Azides; D003432:Cross-Linking Reagents; D007096:Imidoesters; D010797:Phthalimides; D011952:Receptors, Concanavalin A; D013388:Succinimides; D003208:Concanavalin A; C017502:methyl 3-((4-azidophenyl)dithio)propionimidate; C033168:N-((4-azidophenyl)thio)phthalimide; C033169:3-((4-azidophenyl)dithio)propionic N-hydroxysuccinimide