title: Versatile Platform for the Synthesis of Orthogonally Cleavable Heteromultifunctional Cross-Linkers.
abstract: Cleavable and heteromultifunctional cross-linkers have proven critical in a wide range of biological applications. Traditional approaches for synthesizing these linkers suffer from various synthetic and functional limitations. In this work, an efficient sequence-defined synthetic methodology, developed for the assembly of oligothioetheramides, was used to address many of these limitations. Four heterotrifunctional cross-linkers with up to two orthogonal internal cleavage sites were synthesized. These linkers were conjugated to a pair of fluorophores that undergo Förster resonance energy transfer (FRET) and a model protein-human transferrin. Orthogonal bond cleavage was validated by mass spectrometry, fluorescent gel electrophoresis, and confocal microscopy. These studies demonstrate the versatility and biological utility of oligothioetheramides as a new class of multifunctional chemical cross-linkers and biologically relevant fluorescent probes.
authors.affiliation: Robert Frederick Smith School of Chemical and Biomolecular Engineering, Cornell University , Ithaca, New York 14853, United States.
authors.forename: Michelle R
authors.initials: MR
authors.lastname: Sorkin
authors.affiliation: Robert Frederick Smith School of Chemical and Biomolecular Engineering, Cornell University , Ithaca, New York 14853, United States.
authors.forename: Joshua A
authors.initials: JA
authors.lastname: Walker
authors.affiliation: Robert Frederick Smith School of Chemical and Biomolecular Engineering, Cornell University , Ithaca, New York 14853, United States.
authors.forename: Joseph S
authors.initials: JS
authors.lastname: Brown
authors.affiliation: Robert Frederick Smith School of Chemical and Biomolecular Engineering, Cornell University , Ithaca, New York 14853, United States.
authors.forename: Christopher A
authors.identifier: 0000-0003-2654-018X
authors.initials: CA
authors.lastname: Alabi
mesh_terms: D000577:Amides / Q000138:chemical synthesis / Q000737:chemistry*; D001896:Boron Compounds / Q000737:chemistry; D003432:Cross-Linking Reagents / Q000138:chemical synthesis / Q000737:chemistry*; D004987:Ethers / Q000138:chemical synthesis / Q000737:chemistry; D031541:Fluorescence Resonance Energy Transfer; D005456:Fluorescent Dyes / Q000737:chemistry*; D006367:HeLa Cells; D006801:Humans; D018613:Microscopy, Confocal; D008958:Models, Molecular; D014168:Transferrin / Q000737:chemistry*
chemical_list: C095489:4,4-difluoro-4-bora-3a,4a-diaza-s-indacene; D000577:Amides; D001896:Boron Compounds; D003432:Cross-Linking Reagents; D004987:Ethers; D005456:Fluorescent Dyes; D014168:Transferrin