Correlating single-molecule rupture mechanics with cell population adhesion by yeast display

Area of Science:

• Biophysics
• Cell Biology
• Biochemistry

Background:

• Understanding cell adhesion strength and receptor-ligand mechanics is crucial for various biological processes.
• Current methods often lack the throughput or precision to correlate population-level adhesion with single-molecule behavior.

Purpose of the Study:

• To develop and validate a high-throughput yeast surface display method for comparing population-level cell adhesion strength with single-molecule receptor-ligand rupture mechanics.
• To establish correlations between single-molecule force spectroscopy (SMFS) measurements and cell population adhesion strength.

Main Methods:

• Yeast surface display of monomeric streptavidin (mSA) variants.
• High-throughput yeast adhesion assay using a spinning disk to apply shear stress.
• Atomic force microscope single-molecule force spectroscopy (SMFS) to measure rupture forces.
• Quantification of midpoint detachment shear stress for cell populations.

Main Results:

• Established a direct correlation between single-molecule rupture force distributions and cell population adhesion strength.
• Identified key factors for successful correlation, including covalent receptor attachment, controlled pulling geometry, and minimized non-specific adhesion.
• Demonstrated that spinning disk assays can be correlated with SMFS for screening mechanical strength of receptor-ligand complexes.

Conclusions:

• The developed yeast display method enables simultaneous assessment of population-level adhesion and single-molecule mechanics.
• This approach accelerates the study of mechanostable receptor-ligand complexes and receptor-mediated cell adhesion.
• Workflow enhancements facilitate research on the mechanical properties of molecular interactions.