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A giant liposome for single-molecule observation of conformational changes in membrane proteins

Y. Onoue ; T. Suzuki ; M. Davidson ; M. Karlsson ; Owe Orwar (Institutionen för kemi- och bioteknik, Fysikalisk kemi) ; M. Yoshida ; K. Kinosita
Biochimica Et Biophysica Acta-Biomembranes (0005-2736). Vol. 1788 (2009), 6, p. 1332-1340.
[Artikel, refereegranskad vetenskaplig]

We present an experimental system that allows visualization of conformational changes in membrane proteins at the single-molecule level. The target membrane protein is reconstituted in a giant liposome for independent control of the aqueous environments on the two sides of the membrane. For direct observation of conformational changes, an extra-liposomal site(s) of the target protein is bound to a glass surface, and a probe that is easily visible under a microscope, such as a micron-sized plastic bead, is attached to another site on the intra-liposomal side. A conformational change, or an angular motion in the tiny protein molecule, would manifest as a visible motion of the probe. The attachment of the protein on the glass surface also immobilizes the liposome, greatly facilitating its manipulation such as the probe injection. As a model system, we reconstituted ATP synthase (FOF1) in liposomes tens of mu m in size, attached the protein specifically to a glass surface, and demonstrated its ATP-driven rotation in the membrane through the motion of a submicron bead. (c) 2009 Elsevier B.V. All rights reserved.

Nyckelord: Single-molecule, Giant liposome, Membrane protein, Conformational, change, ATP synthase, mitochondrial adenosine-triphosphatase, resonance energy-transfer, c-subunit ring, atp synthase, escherichia-coli, unilamellar vesicles, stepwise rotation, kinetic mechanism, flagellar motor, channel



Denna post skapades 2010-01-14. Senast ändrad 2010-04-30.
CPL Pubid: 107240

 

Institutioner (Chalmers)

Institutionen för kemi- och bioteknik, Fysikalisk kemi (2005-2014)

Ämnesområden

Kemi
Fysikalisk kemi

Chalmers infrastruktur