Abstract Uzun
Functional reconstitution of plant P3 ATPase into giant unilamellar vesicles
Membrane transporters are essential for numerous biological processes by controlling the movement of ions and molecules across cell membranes. However, dissecting their molecular dynamics in complex cellular environments presents significant challenges. Reconstitution of membrane transporters in model systems offers a powerful solution. This study focused on the reconstitution conditions suitable for a plant P3 ATPase and compatible with various giant unilamellar vesicle generation techniques. Among the methods evaluted for GUV formation, including electroformation, gel-assisted
formation, and charge-mediated fusion, only the gel-assisted approach successfully generated P3 ATPase-containing giant unilamellar vesicles while preserving the pump activity. These results underscore the importance of carefully managing the reconstitution conditions, including the presence of ions, and selecting the appropriate lipid composition to enhance the stability and activity of P3 ATPase in proteoliposomes. Addressing these factors is essential for the successful formation and functional analysis of P3 ATPase and other Ptype ATPases in experimental settings.
Huriye D. Uzun, Ekaterina Malysenko, Bo H. Justesen and Thomas Günther Pomorski; Department of Molecular Biochemistry, Faculty of Chemistry and Biochemistry, Ruhr University Bochum, Bochum, Germany