AFM visualization of mobile influenza a M2 molecules in planar bilayers

Travis Hughes; Bradley Strongin; Fei Philip Gao; Viksita Vijayvergiya; David D. Busath; Robert C. Davis

doi:10.1529/biophysj.103.036111

AFM visualization of mobile influenza a M2 molecules in planar bilayers

Travis Hughes
, Bradley Strongin
, Fei Philip Gao
, Viksita Vijayvergiya
, David D. Busath
, Robert C. Davis

Research output: Contribution to journal › Article › peer-review

16 Scopus citations

Abstract

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 × 10^-14 cm²/s. Eighty-two percent of observable particles were mobile on the observable timescale (D > 6 × 10 ^-15 cm²/s). Protein-protein interactions were also observed directly.

Original language	English
Pages (from-to)	311-322
Number of pages	12
Journal	Biophysical Journal
Volume	87
Issue number	1
DOIs	https://doi.org/10.1529/biophysj.103.036111
State	Published - Jul 2004

Funding

This work was supported in part by National Institutes of Health grant AI23007.

Funder number
R01AI023007

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10.1529/biophysj.103.036111

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title = "AFM visualization of mobile influenza a M2 molecules in planar bilayers",

abstract = "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 × 10-14 cm2/s. Eighty-two percent of observable particles were mobile on the observable timescale (D > 6 × 10 -15 cm2/s). Protein-protein interactions were also observed directly.",

author = "Travis Hughes and Bradley Strongin and Gao, \{Fei Philip\} and Viksita Vijayvergiya and Busath, \{David D.\} and Davis, \{Robert C.\}",

note = "Funding Information: This work was supported in part by National Institutes of Health grant AI23007.",

year = "2004",

month = jul,

doi = "10.1529/biophysj.103.036111",

language = "English",

volume = "87",

pages = "311--322",

journal = "Biophysical Journal",

issn = "0006-3495",

publisher = "Elsevier Inc.",

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T1 - AFM visualization of mobile influenza a M2 molecules in planar bilayers

AU - Hughes, Travis

AU - Strongin, Bradley

AU - Gao, Fei Philip

AU - Vijayvergiya, Viksita

AU - Busath, David D.

AU - Davis, Robert C.

N1 - Funding Information: This work was supported in part by National Institutes of Health grant AI23007.

PY - 2004/7

Y1 - 2004/7

N2 - 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 × 10-14 cm2/s. Eighty-two percent of observable particles were mobile on the observable timescale (D > 6 × 10 -15 cm2/s). Protein-protein interactions were also observed directly.

AB - 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 × 10-14 cm2/s. Eighty-two percent of observable particles were mobile on the observable timescale (D > 6 × 10 -15 cm2/s). Protein-protein interactions were also observed directly.

UR - https://www.scopus.com/pages/publications/3042815787

U2 - 10.1529/biophysj.103.036111

DO - 10.1529/biophysj.103.036111

M3 - Article

C2 - 15240466

AN - SCOPUS:3042815787

SN - 0006-3495

VL - 87

SP - 311

EP - 322

JO - Biophysical Journal

JF - Biophysical Journal

IS - 1

ER -

AFM visualization of mobile influenza a M2 molecules in planar bilayers

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