Cell surface display of a protein based on a tail-anchored membrane protein

Area of Science:

• Biotechnology
• Bioengineering
• Molecular Biology

Background:

• Cell surface display is crucial for protein-based biotechnology and bioengineering.
• Current methods often use type I transmembrane proteins for anchoring, limiting display orientation.
• Tail-anchored (TA) proteins offer an alternative anchoring strategy with C-terminal surface exposure.

Purpose of the Study:

• To develop an efficient cell surface display system utilizing the transmembrane domain (TMD) of a TA protein.
• To enable N-terminal fusion of target proteins to the TMD, preserving C-terminal activity.
• To overcome limitations of existing protein display technologies.

Main Methods:

• Constructed fusion proteins using the TMD of the TA protein emerin.
• Engineered truncated forms of emerin for optimized surface display.
• Expressed fusion constructs in mammalian cells to assess translocation and display.

Main Results:

• A truncated emerin construct (lacking N-terminal 210 residues) served as an effective TMD anchor.
• Efficient translocation of fusion proteins to the plasma membrane was achieved.
• Successful display of target proteins on the cell surface was confirmed.

Conclusions:

• The developed system provides an effective method for cell surface protein display.
• Utilizing TA protein TMDs enhances protein display applicability, especially for proteins with critical C-terminal functions.
• This technology advances cell surface display capabilities in biotechnology and bioengineering.