Enzyme Fueled Dissipative Self-assembly of Guanine Functionalized Molecules and Their Cellular Behaviour

Area of Science:

• Biochemistry
• Materials Science
• Nanotechnology

Background:

• Esterase lipase enzymes typically hydrolyze esters in aqueous solutions.
• G-quadruplex structures are nucleic acid secondary structures with potential in self-assembly.
• Understanding enzyme-driven self-assembly is crucial for biomaterial development.

Purpose of the Study:

• To investigate the effect of G-quadruplex self-assembly on reverse hydrolysis equilibrium.
• To synthesize and characterize novel peptide-lipid-conjugated guanine base molecules for self-assembly.
• To explore the potential of enzyme-fueled dissipative self-assembled structures as biomaterials.

Main Methods:

• Synthesis of five peptide-lipid-conjugated guanine base functionalized molecules.
• Confirmation of G-quadruplex structure using spectroscopic techniques (XRD, ThT assay, CD).
• Analysis of self-assembled structures (nanofibers within spheres) using CLSM, FE-SEM, and HR-TEM.
• Biocompatibility assessment using MTT assay on HEK 293 and McCoy cell lines.
• Evaluation of wound repair potential via scratch assay.

Main Results:

• NAC5 molecule successfully self-assembled into a G-quadruplex structure.
• Enzyme (lipase) fueled self-assembly resulted in nanofibers enclosed within spheres.
• The synthesized G-quadruplex hydrogel exhibited good biocompatibility and low cytotoxicity.
• The NAC5 hydrogel demonstrated potential for wound repair applications through cell migration.

Conclusions:

• G-quadruplex self-assembly can effectively shift hydrolysis equilibrium in reverse.
• Enzyme-fueled dissipative self-assembly offers a novel route to create functional nanostructures.
• The developed NAC5 hydrogel is a promising biocompatible biomaterial for protocell applications and wound healing.