Construction of a stable functional ribonucleopeptide complex by the covalent linking method.
We describe here a novel strategy to create a stable functional ribonucleopeptide (RNP) complex by the covalent linking method. Adenosine-5'-triphosphate (ATP)-binding RNP receptors were selected from the RNP library by in vitro selection. The RNA subunit of RNP is utilized to construct a ligand-binding cavity, while the peptide subunit can be functionalized independently. By introducing a fluorophore at the N-terminus of the Rev peptide subunit, the ATP-binding RNP receptor is successfully converted to a noncovalent complex of ATP-responsive fluorescent RNP sensor. Such a noncovalent RNP sensor could be covalently linked by the tethering the RNA to the fluorophore-labeled peptide subunit to form a stable RNP sensor without losing the original function.
Fukuda
Masatora
M
Pioneering Research Unit for Next Generation, Kyoto university, Uji, Kyoto 611-0011, Japan.
Nakano
Shun
S
Tainaka
Kazuki
K
Fujieda
Nobutaka
N
Morii
Takashi
T
D000255:Adenosine Triphosphate / Q000032:analysis; D005456:Fluorescent Dyes / Q000737:chemistry*; D008024:Ligands; D020411:Oligonucleotide Array Sequence Analysis; D010452:Peptide Biosynthesis; D010455:Peptides / Q000737:chemistry*; D012313:RNA / Q000096:biosynthesis / Q000737:chemistry*
D005456:Fluorescent Dyes; D008024:Ligands; D010455:Peptides; D012313:RNA; D000255:Adenosine Triphosphate