| 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 |
3.412
|
| Times Cited |
19
|
|
WOS Category
|
CHEMISTRY, ORGANIC
|
| Resource |
| DNA material |
ETR103 (RDB01198) |
