Bottom-Up Approach to Explore Alpha-Amylase Assisted Membrane Remodelling

TL;DR

This study investigates how alpha-amylase interacts with lipid membranes, altering their mechanical properties and inducing morphological changes, with implications for membrane remodeling processes.

Contribution

It demonstrates that alpha-amylase can modulate membrane rigidity and curvature, revealing a novel role in membrane remodeling beyond its enzymatic activity.

Findings

Alpha-amylase increases membrane bending rigidity at low concentrations.

Higher concentrations of alpha-amylase induce membrane curvature and morphological changes.

At very high concentrations, membranes collapse into highly convoluted structures.

Abstract

Soluble alpha-amylases play an important role in the catabolism of polysaccharides. In this work, we show that the enzyme can interact with the lipid membrane and further alter its mechanical properties. Vesicle fluctuation spectroscopy is used for quantitative measurement of the membrane bending rigidity of phosphatidylcholines lipid vesicles from the shape fluctuation based on the whole contour of Giant Unilamellar Vesicles (GUVs). The bending rigidity of the lipid vesicles of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine in water increases significantly with the presence of 0.14 micromolar alpha-amylase in the exterior solution. However, as the concentration increases above 1 micromolar, the bending rigidity decreases but remains higher than estimated without the protein. Contact between the alpha-amylase in the outer solution and the outer leaflet leads to spontaneous membrane curvature and the corresponding morphological changes of the GUVs. The presence of outbuds directly demonstrates that AA has a preferable interaction with the membrane, giving a positive spontaneous curvature of $C_0 \leq 0.05 \ \mu \rm{m}^{-1}$ at $18 \ \mu$M $(\approx$ 1 mg/ml) of AA concentration. Above 1 mg/ml of AA concentration the shape of GUVs collapse completely suggesting a highly convoluted state.