RRC ID 3805
Author Takeda S, Tsukiji S, Ueda H, Nagamune T.
Title Covalent split protein fragment-DNA hybrids generated through N-terminus-specific modification of proteins by oligonucleotides.
Journal Org Biomol Chem
Abstract Semisynthetic protein-DNA hybrid molecules have recently attracted much attention as valuable tools for bioanalytical chemistry and nanobiotechnology. Here we describe a synthetic method for conjugating oligonucleotides to the N-terminus of recombinant proteins. Our strategy involves the conversion of amine-terminated oligonucleotides to thioester-functionalized oligonucleotides by using a bifunctional reagent bearing an N-hydroxysuccinimide ester and benzyl thioester group, followed by native chemical ligation with proteins containing an N-terminal cysteine. We applied this technique to construct split luciferase fragment-DNA hybrid systems in which the catalytic activity of split luciferase is restored by the re-assembly of each fragment through a specific DNA-protein or DNA-DNA interaction. Split protein fragment-DNA hybrids will offer new opportunities to explore the potential of protein-DNA conjugates for various applications.
Volume 6(12)
Pages 2187-94
Published 2008-6-21
DOI 10.1039/b720013g
PMID 18528581
MeSH Base Sequence Chromatography, High Pressure Liquid Oligonucleotides / chemistry* Proteins / chemistry* Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
IF 3.49
Times Cited 19
DNA material ETR103 (RDB01198)