A novel method of AquaporinZ incorporation via binary-lipid Langmuir monolayers

[Display omitted] ► The incorporation of Histidine-tagged AquaporinZ in a lipid bilayer is investigated with Langmuir–Blodgett technology for the first time. ► Detergent removal in the Langmuir–Blodgett system is studied. ► AquaporinZ and detergent interaction during protein insertion is elaborated and a protein incorporation mechanism is proposed. In this work, a new approach of incorporating transmembrane protein AquaporinZ (AqpZ) into a lipid bilayer has been developed with the aid of the Langmuir–Blodgett technique. The binary-lipid monolayer for AqpZ incorporation is composed of (1) gel-phase lipids resistant to detergent dissolution and (2) nickel-chelating lipids that can attach the histidine-tagged AqpZ from the subphase. Upon removal of subphase detergent with BioBeads, the incorporation is achieved by transferring the AqpZ-associated binary-lipid monolayer onto a preformed pure binary-lipid monolayer using the Langmuir–Schaefer deposition method. AFM images show an indication of AqpZ incorporation in the bilayer. Furthermore, it is also shown that BioBeads can remove a significant amount of detergent in the subphase and lipid film integrity is restored after detergent removal. The detergent removal rate is correlated to BioBeads amount and subphase circulation. The new approach of AqpZ reconstitution revealed in this work could potentially be applied in biomimetic membrane formation for water purification applications.