Microtubule self-healing and defect creation investigated by in-line force measurements during high-speed atomic...

Area of Science:

• Biophysics
• Cell Biology
• Materials Science

Background:

• Microtubules are essential eukaryotic biopolymers involved in cell division, migration, and transport.
• Mechanical stress can cause defects in microtubules, altering their function and interactions with associated proteins.
• Understanding these defects is crucial for comprehending fundamental cellular processes.

Purpose of the Study:

• To develop a method for controlled defect creation in microtubules.
• To simultaneously gather mechanical information and image dynamic processes at the molecular level.
• To quantitatively assess the mechanical properties and bonding energy of tubulin dimers.

Main Methods:

• Utilized high-speed atomic force microscopy (HS-AFM).
• Developed and applied an 'in-line force curve mode' for real-time defect induction and monitoring.
• Collected force curve data during defect formation.

Main Results:

• Successfully created defects in microtubules at the tubulin dimer level.
• Monitored dynamic processes occurring around induced defects in real-time.
• Obtained quantitative mechanical data, enabling estimation of tubulin dimer bonding energy.

Conclusions:

• The in-line force curve mode of HS-AFM is effective for studying microtubule mechanics and defect dynamics.
• This method provides novel insights into the relationship between microtubule structure, mechanical properties, and function.
• The technique facilitates a deeper understanding of molecular interactions within biopolymers.