title: Efficient Synthesis of Peptide and Protein Functionalized Pyrrole-Imidazole Polyamides Using Native Chemical Ligation.
abstract: The advancement of DNA-based bionanotechnology requires efficient strategies to functionalize DNA nanostructures in a specific manner with other biomolecules, most importantly peptides and proteins. Common DNA-functionalization methods rely on laborious and covalent conjugation between DNA and proteins or peptides. Pyrrole-imidazole (Py-Im) polyamides, based on natural minor groove DNA-binding small molecules, can bind to DNA in a sequence specific fashion. In this study, we explore the use of Py-Im polyamides for addressing proteins and peptides to DNA in a sequence specific and non-covalent manner. A generic synthetic approach based on native chemical ligation was established that allows efficient conjugation of both peptides and recombinant proteins to Py-Im polyamides. The effect of Py-Im polyamide conjugation on DNA binding was investigated by Surface Plasmon Resonance (SPR). Although the synthesis of different protein-Py-Im-polyamide conjugates was successful, attenuation of DNA affinity was observed, in particular for the protein-Py-Im-polyamide conjugates. The practical use of protein-Py-Im-polyamide conjugates for addressing DNA structures in an orthogonal but non-covalent manner, therefore, remains to be established.
authors.affiliation: Laboratory of Chemical Biology and Institute for Complex Molecular Systems, Eindhoven University of Technology, Den Dolech 2, 5600 MB Eindhoven, The Netherlands. b.m.g.janssen@tue.nl.
authors.forename: Brian M G
authors.initials: BM
authors.lastname: Janssen
authors.affiliation: Laboratory of Chemical Biology and Institute for Complex Molecular Systems, Eindhoven University of Technology, Den Dolech 2, 5600 MB Eindhoven, The Netherlands. svenvanommeren@gmail.com.
authors.forename: Sven P F I
authors.initials: SP
authors.lastname: van Ommeren
authors.affiliation: Laboratory of Chemical Biology and Institute for Complex Molecular Systems, Eindhoven University of Technology, Den Dolech 2, 5600 MB Eindhoven, The Netherlands. m.merkx@tue.nl.
authors.forename: Maarten
authors.initials: M
authors.lastname: Merkx
mesh_terms: D004247:DNA / Q000737:chemistry* / Q000378:metabolism; D007093:Imidazoles / Q000737:chemistry; D015394:Molecular Structure; D009757:Nylons / Q000138:chemical synthesis* / Q000737:chemistry; D010455:Peptides / Q000737:chemistry / Q000378:metabolism*; D011758:Pyrroles / Q000737:chemistry; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism*; D020349:Surface Plasmon Resonance
chemical_list: D007093:Imidazoles; D009757:Nylons; D010455:Peptides; D011758:Pyrroles; D011994:Recombinant Proteins; C029899:imidazole; D004247:DNA
keywords: DNA-functionalization; DNA-nanotechnology; native chemical ligation; pyrrole-imidazole polyamides; surface plasmon resonance