Recognition-then-Reaction Enables Site-Selective Bioconjugation to Proteins on Live-Cell Surfaces

Area of Science:

• Bioconjugation Chemistry
• Molecular Biology
• Biotechnology

Background:

• Site-selective protein modification is crucial for protein functionalization but often requires laborious genetic engineering.
• Existing methods are complex, time-consuming, and technically challenging.

Purpose of the Study:

• To develop a novel, efficient, and site-selective method for protein modification using aptamers.
• To enable single-step protein/DNA conjugation under mild conditions.

Main Methods:

• Development of an aptamer-based recognition-then-reaction strategy.
• Application of the method to model proteins like human thrombin, PDGF-BB, Avidin, and His-tagged proteins.
• Utilizing cell-SELEX to obtain aptamers for live-cell applications.

Main Results:

• Achieved outstanding selectivity and efficiency in site-selective protein/DNA conjugation.
• Demonstrated excellent results on various model proteins under mild reaction conditions.
• Successfully enabled real-time modification of live-cell membrane proteins without pre-treatment.

Conclusions:

• The aptamer-based recognition-then-reaction method offers a simple, highly selective, and efficient approach for protein modification.
• This technique facilitates protein tagging in complex biological environments and in real-time on live cells.
• The method overcomes limitations of traditional protein engineering approaches for functionalization.