Domain-Targeted Membrane Partitioning of Specific Proteins with DNA Nanodevices

Area of Science:

• Cell Biology
• Biotechnology
• Nanotechnology

Background:

• Cell membranes feature complex lipid and protein domains crucial for cellular functions.
• Manipulating membrane protein localization without genetic engineering is vital but challenging for understanding cellular processes.

Purpose of the Study:

• To develop novel DNA nanodevices for targeted manipulation of membrane protein domains.
• To investigate the functional consequences of altering protein localization in lipid-ordered (Lo) and lipid-disorder (Ld) domains on live cells.

Main Methods:

• Conjugation of cholesterols or tocopherols to DNA tetrahedrons to create nanodevices targeting Lo and Ld domains.
• Incorporation of protein-recognition ligands (aptamers, antibodies) via toehold-mediated strand displacement for protein translocation.
• Utilizing PTK7 and CD45 as model proteins to demonstrate nanodevice efficacy in regulating cell migration and T cell activation.

Main Results:

• Demonstrated that PTK7 accumulation in Lo domains promotes tumor cell migration, while sequestration in Ld domains inhibits it.
• Showcased the modularity of DNA nanodevices in regulating T cell activation by manipulating CD45 translocation.
• Established a method for dynamic control of protein localization within distinct membrane domains on live cells.

Conclusions:

• Developed versatile DNA nanodevices capable of precise manipulation of membrane protein localization.
• Provided insights into the role of PTK7 and CD45 localization in cell migration and T cell activation.
• Opened new avenues for studying membrane structure, molecular interactions, and cell signaling pathways.