Solubilization of Membrane Proteins using designed protein WRAPS.

Area of Science:

• Biochemistry
• Structural Biology
• Protein Engineering

Background:

• Integral membrane proteins present significant challenges for therapeutic and vaccine development due to their hydrophobic nature, complicating production and structural analysis.
• Current methods often rely on detergents, which can interfere with protein function and stability.

Purpose of the Study:

• To introduce a general deep learning-based design approach for creating water-soluble versions of native membrane proteins.
• To demonstrate that these engineered proteins (WRAPs) retain native function, sequence, and fold while being stable and detergent-free.

Main Methods:

• Utilized a deep learning approach to design genetically encoded de novo proteins called WRAPs (Water-soluble RFdiffused Amphipathic Proteins).
• Engineered WRAPs to surround the hydrophobic surfaces of both beta-barrel and helical membrane proteins, rendering them soluble.
• Applied the method to design soluble versions of four Treponema pallidum outer membrane beta barrels, potential syphilis vaccine antigens.

Main Results:

• Designed and validated WRAPs for both beta-barrel and helical membrane proteins, achieving water solubility without detergents.
• Demonstrated that WRAP-solubilized proteins maintain their original binding and enzymatic functions and exhibit enhanced stability.
• Generated soluble antigens for syphilis vaccine development and obtained a cryo-EM map of a WRAPed protein consistent with the design.

Conclusions:

• WRAP technology offers a versatile platform for overcoming the challenges associated with integral membrane proteins.
• This approach facilitates biochemical and structural characterization, accelerates therapeutic discovery through screening of soluble targets, and aids in developing novel vaccines.