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Biophysical Journal 87:311-322 (2004)
© 2004 The Biophysical Society

AFM Visualization of Mobile Influenza A M2 Molecules in Planar Bilayers

Travis Hughes *, Bradley Strongin *, Fei Philip Gao {dagger}, Viksita Vijayvergiya {ddagger}, David D. Busath {ddagger} and Robert C. Davis *

* Department of Physics and Astronomy, and {ddagger} Department of Physiology and Developmental Biology, Brigham Young University, Provo, Utah; and {dagger} National High Magnetic Field Laboratory and Department of Chemistry and Biochemistry, Florida State University, Tallahassee, Florida

Correspondence: Address reprint requests to Travis Hughes, University of Colorado, MCDB, 347 UCB, Boulder, CO 80309-0347. Tel.: 303-786-1007; Fax: 303-492-7744; E-mail: Travis.Hughes{at}colorado.edu.

We report the observation of influenza A M2 (M2) incorporated in a dipalmitoylphosphatidylcholine (DPPC) supported planar bilayer on mica, formed by use of a modified vesicle fusion method from proteoliposomes and visualized with contact mode atomic force microscopy. Incubation of proteoliposomes in a hyperosmotic solution and increased DPPC/M2 weight ratios improved supported planar bilayer formation by M2/DPPC proteoliposomes. M2's extra-bilayer domains were observed as particles estimated to protrude 1–1.5 nm above the bilayer surface and <4 nm in diameter. Particle density was 5–18% of the nominal tetramer density. Movement of observable M2 particles was independent of the probe tip. The mean lateral diffusion coefficient (D) of M2 was 4.4 ± 1.0 x 10–14 cm2/s. Eighty-two percent of observable particles were mobile on the observable timescale (D > 6 x 10–15 cm2/s). Protein-protein interactions were also observed directly.






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Copyright © 2004 by the Biophysical Society.