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

Yasuhiro Onoue, Toshiharu Suzuki, Max Davidson, Mattias Karlsson, Owe Orwar, Masasuke Yoshida, Kazuhiko Kinosita

    Research output: Contribution to journalArticle

    7 Citations (Scopus)

    Abstract

    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 μm 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.

    Original languageEnglish
    Pages (from-to)1332-1340
    Number of pages9
    JournalBiochimica et Biophysica Acta - Biomembranes
    Volume1788
    Issue number6
    DOIs
    Publication statusPublished - 2009 Jun

    Fingerprint

    Liposomes
    Membrane Proteins
    Observation
    Glass
    Molecules
    Proteins
    Adenosine Triphosphate
    Membranes
    Plastics
    Microscopes
    Visualization
    Injections

    Keywords

    • ATP synthase
    • Conformational change
    • Giant liposome
    • Membrane protein
    • Single-molecule

    ASJC Scopus subject areas

    • Biochemistry
    • Cell Biology
    • Biophysics

    Cite this

    Onoue, Y., Suzuki, T., Davidson, M., Karlsson, M., Orwar, O., Yoshida, M., & Kinosita, K. (2009). A giant liposome for single-molecule observation of conformational changes in membrane proteins. Biochimica et Biophysica Acta - Biomembranes, 1788(6), 1332-1340. https://doi.org/10.1016/j.bbamem.2009.01.015

    A giant liposome for single-molecule observation of conformational changes in membrane proteins. / Onoue, Yasuhiro; Suzuki, Toshiharu; Davidson, Max; Karlsson, Mattias; Orwar, Owe; Yoshida, Masasuke; Kinosita, Kazuhiko.

    In: Biochimica et Biophysica Acta - Biomembranes, Vol. 1788, No. 6, 06.2009, p. 1332-1340.

    Research output: Contribution to journalArticle

    Onoue, Y, Suzuki, T, Davidson, M, Karlsson, M, Orwar, O, Yoshida, M & Kinosita, K 2009, 'A giant liposome for single-molecule observation of conformational changes in membrane proteins', Biochimica et Biophysica Acta - Biomembranes, vol. 1788, no. 6, pp. 1332-1340. https://doi.org/10.1016/j.bbamem.2009.01.015
    Onoue, Yasuhiro ; Suzuki, Toshiharu ; Davidson, Max ; Karlsson, Mattias ; Orwar, Owe ; Yoshida, Masasuke ; Kinosita, Kazuhiko. / A giant liposome for single-molecule observation of conformational changes in membrane proteins. In: Biochimica et Biophysica Acta - Biomembranes. 2009 ; Vol. 1788, No. 6. pp. 1332-1340.
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