The stability and functionality of chemically crosslinked microtubules.

Area of Science:

• Biochemistry
• Materials Science
• Nanotechnology

Background:

• Microtubules (MTs) are crucial for intracellular transport and are being explored as shuttles for nanomaterials.
• Stabilizing MTs is essential for their use in artificial active transport systems.
• Existing stabilization methods, like taxol, have limitations.

Purpose of the Study:

• To explore bifunctional crosslinking agents for stabilizing MT shuttles.
• To investigate how different crosslinking agent attributes affect MT stability and functionality.
• To compare amine-targeting vs. cysteine-targeting crosslinkers.

Main Methods:

• Utilized various bifunctional crosslinking agents targeting amine and cysteine residues on tubulin.
• Assessed the structural and functional stability of modified microtubules (CLMTs).
• Evaluated CLMTs' performance in active transport assays with kinesin-coated surfaces.

Main Results:

• Amine-targeting crosslinkers formed CLMTs with up to four times longer structural and functional lifetimes than taxol.
• CLMTs demonstrated stability at low temperatures (-10°C) and resistance to Ca2+ depolymerization.
• Cysteine-targeting crosslinkers led to MT depolymerization, likely by inhibiting GTP binding.

Conclusions:

• Bifunctional crosslinking agents, particularly those targeting amines, offer superior stabilization for MTs in active transport applications.
• CLMTs exhibit enhanced robustness and maintained functionality for nanomaterial delivery systems.
• Crosslinker design, including terminal group chemistry and chain length, critically influences MT stabilization efficacy.