Abstract
The function of any given biological membrane is determined largely by the specific set of integral membrane proteins embedded in it, and the peripheral membrane proteins attached to the membrane surface. The activity of these proteins, in turn, can be modulated by the phospholipid composition of the membrane. The reconstitution of membrane proteins into a model membrane allows investigation of individual features and activities of a given cell membrane component. However, the activity of membrane proteins is often difficult to sustain following reconstitution, since the composition of the model phospholipid bilayer differs from that of the native cell membrane. This review will discuss the reconstitution of membrane protein activities in four different types of model membrane—monolayers, supported lipid bilayers, liposomes and nanodiscs, comparing their advantages in membrane protein reconstitution. Variation in the surrounding model environments for these four different types of membrane layer can affect the three-dimensional structure of reconstituted proteins and may possibly lead to loss of the proteins activity. We also discuss examples where the same membrane proteins have been successfully reconstituted into two or more model membrane systems with comparison of the observed activity in each system. Understanding of the behavioral changes for proteins in model membrane systems after membrane reconstitution is often a prerequisite to protein research. It is essential to find better solutions for retaining membrane protein activities for measurement and characterization in vitro.
Highlights
The cell membrane separates intracellular components from the outside environment and is constituted by various phospholipids, cholesterol, glycolipids and proteins
The interfacial recognition and adsorption of phospholipases A2 (PLA2) and phospholipases C (PLC) to the phospholipid membrane interface are poorly understood. It appears that both PLA2 and PLC are active at the monolayer model membrane, indicating that the kinetics of phospholipid hydrolysis at the air–water interface can be monitored by biophysical characterization techniques in situ such as PM-IRRAS and infrared reflection adsorption spectroscopy [22]
This review summarizes and compares the most up-to-date methods for reconstituting membrane proteins into model membranes
Summary
The cell membrane separates intracellular components from the outside environment and is constituted by various phospholipids, cholesterol, glycolipids and proteins. Other factors affecting the reconstituted membrane protein activity are the chemical properties of the lipid head groups which control membrane hydrophilicity. Both parameters are crucial in stabilizing membrane protein structure. We describe four case studies and will compare the protein activity changes when the membrane proteins are reconstituted into different model membranes. In these case studies, we demonstrate how protein activities are modulated by the lipid environment and discuss how this environment helps to retain protein activities.
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