Protein Labeling and Crosslinking by Covalent Aptamers

Area of Science:

• Biochemistry
• Molecular Biology
• Chemical Biology

Background:

• Traditional aptamers face limitations like nuclease instability and short target engagement times.
• Selective modification of native proteins in their native environment remains a challenge.

Purpose of the Study:

• To develop a novel method for selective protein modification using engineered aptamers.
• To create covalent aptamers capable of stable target engagement and functionalization.

Main Methods:

• Engineered electrophilic covalent aptamers by introducing proximity-driven electrophiles at specific nucleotide sites.
• Utilized thrombin as a model protein to demonstrate the functionality of covalent aptamers.
• Assessed the ability of covalent aptamers to transfer functional handles and crosslink to target proteins.

Main Results:

• Demonstrated selective modification of native proteins in their biological environment.
• Showcased the transfer of functional handles and irreversible crosslinking to target proteins.
• Covalent aptamers exhibited enhanced stability against nucleases and prolonged target engagement.

Conclusions:

• Covalent aptamers represent a versatile new tool for specific protein modification and sensitive detection.
• This approach overcomes common aptamer limitations, offering improved stability and residence time.
• Covalent aptamers enable nuclease-resistant enzyme inhibition and precise protein manipulation.