Nanodisc

Isolation of Monodisperse Nanodisc-Reconstituted Membrane Proteins Using Free Flow Electrophoresis

Bo Højen Justesen,†,∥ Tomas Laursen,‡,∥ Gerhard Weber,§ Anja Thoe Fuglsang,† Birger Lindberg Møller,*,‡ and Thomas Günther Pomorski*,† Section for Transport Biology, Department of Plant and Environmental Sciences, Faculty of Science, University of Copenhagen,
Thorvaldsenvej 40, DK-1871 Frederiksberg C, Denmark ‡Plant Biochemistry Laboratory, Department of Plant and Environmental Sciences, Faculty of Science, University of Copenhagen,Thorvaldsenvej 40, DK-1871 Frederiksberg C, Denmark, FFE Service GmbH, Frankfurter Ring 193a, D-80807 Munich, Germany

*supporting Information

ABSTRACT

Free flow electrophoresis is used for rapid and highrecovery isolation of homogeneous preparations of functionally active membrane proteins inserted into nanodiscs. The approachenables isolation of integral and membrane anchored proteins and is also applicable following introduction of, e.g., fluorescent tags. Preparative separation of membrane protein loaded nanodiscs from empty nanodiscs and protein aggregates results in monodisperse nanodisc preparations ideal for structural and functional characterization using biophysical methods.

Received: January 10, 2013
Accepted: March 4, 2013
Published: March 4, 2013