Construction of a stable functional ribonucleopeptide complex by the covalent linking method.

Area of Science:

• Biochemistry
• Molecular Biology
• Biotechnology

Background:

• Ribonucleopeptide (RNP) complexes are crucial for various cellular processes.
• Developing stable and functional RNP complexes for sensing applications remains a challenge.
• Adenosine triphosphate (ATP)-binding RNP receptors offer potential for molecular sensing.

Purpose of the Study:

• To develop a novel strategy for creating stable, functional ribonucleopeptide (RNP) complexes.
• To engineer an ATP-responsive fluorescent RNP sensor.
• To establish a covalent linking method for RNP sensor stabilization.

Main Methods:

• In vitro selection was used to identify adenosine-5'-triphosphate (ATP)-binding RNP receptors from an RNP library.
• The RNA subunit was engineered to form a ligand-binding cavity.
• The peptide subunit was functionalized with a fluorophore, and covalent linking was employed to stabilize the complex.

Main Results:

• A novel strategy for creating stable functional ribonucleopeptide (RNP) complexes via covalent linking was established.
• ATP-binding RNP receptors were successfully selected and engineered.
• An ATP-responsive fluorescent RNP sensor was developed, demonstrating stability without loss of function.

Conclusions:

• The covalent linking method provides a stable and functional RNP sensor.
• This approach enables the creation of robust molecular sensors for detecting specific ligands like ATP.
• The strategy holds promise for applications in molecular diagnostics and biochemical research.