Deconvoluting the Effect of the Hydrophobic and Hydrophilic Domains of an Amphiphilic Integral Membrane Protein in Lipid Bicontinuous Cubic Mesophases.
Lipidic bicontinuous cubic mesophases with encapsulated amphiphilic proteins are widely used in a range of biological and biomedical applications, including in meso crystallization, as drug delivery vehicles for therapeutic proteins, and as biosensors and biofuel cells. However, the effect of amphiphilic protein encapsulation on the cubic phase nanostructure is not well-understood. In this study, we illustrate the effect of incorporating the bacterial amphiphilic membrane protein Ag43, and its individual hydrophobic β(43) and hydrophilic α(43) domains, in bicontinuous cubic mesophases. For the monoolein, monoalmitolein, and phytantriol cubic phases with and without 8% w/w cholesterol, the effect of the full length amphiphilic protein Ag43 on the cubic phase nanostructure was more significant than the sum of the individual hydrophobic β(43) and hydrophilic α(43) domains. Several factors were found to potentially influence the impact of the hydrophobic β(43) domain on the cubic phase internal nanostructure. These include the size of the hydrophobic β(43) domain relative to the thickness of the lipid bilayer, as well as its charge and diameter. The size of the hydrophilic α(43) domain relative to the water channel radius of the cubic mesophase was also found to be important. The secondary structure of the Ag43 proteins was affected by the hydrophobic thickness and physicochemical properties of the lipid bilayer and the water channel diameter of the cubic phase. Such structural changes may be small but could potentially affect membrane protein function.
CSIRO Manufacturing Flagship , Clayton, Victoria 3169, Australia.
Leonie
L
van 't Hag
Hsin-Hui
HH
Shen
Jingxiong
J
Lu
Australian Synchrotron , Clayton, Victoria 3168, Australia.
Adrian M
AM
Hawley
Sally L
SL
Gras
CSIRO Manufacturing Flagship , Clayton, Victoria 3169, Australia.|School of Applied Sciences, College of Science, Engineering and Health, RMIT University , Melbourne, Victoria 3001, Australia.
Calum J
CJ
Drummond
School of Applied Sciences, College of Science, Engineering and Health, RMIT University , Melbourne, Victoria 3001, Australia.
Charlotte E
CE
Conn
D002942:Circular Dichroism; D057927:Hydrophobic and Hydrophilic Interactions; D008055:Lipids / Q000737:chemistry*; D008565:Membrane Proteins / Q000737:chemistry*
D008055:Lipids; D008565:Membrane Proteins