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Thermal Fluctuations in Amphipol A8-35 Particles: A Neutron Scattering and Molecular Dynamics Study

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Abstract

Amphipols are a class of polymeric surfactants that can stabilize membrane proteins in aqueous solutions as compared to detergents. A8-35, the best-characterized amphipol to date, is composed of a polyacrylate backbone with ~35 % of the carboxylates free, ~25 % grafted with octyl side-chains, and ~40 % with isopropyl ones. In aqueous solutions, A8-35 self-organizes into globular particles with a molecular mass of ~40 kDa. The thermal dynamics of A8-35 particles was measured by neutron scattering in the 10-picosecond, 18-picosecond, and 1-nanosecond time-scales on natural abundance and deuterium-labeled molecules, which permitted to separate backbone and side-chain motions. A parallel analysis was performed on molecular dynamics trajectories (Perlmutter et al., Langmuir 27:10523–10537, 2011). Experimental results and simulations converge, from their respective time-scales, to show that A8-35 particles feature a more fluid hydrophobic core, predominantly containing the octyl chains, and a more rigid solvent-exposed surface, made up predominantly of the hydrophilic polymer backbone. The fluidity of the core is comparable to that of the lipid environment around proteins in the center of biological membranes, as also measured by neutron scattering. The biological activity of proteins depends sensitively on molecular dynamics, which itself is strongly dependent on the immediate macromolecular environment. In this context, the characterization of A8-35 particle dynamics constitutes a step toward understanding the effect of amphipols on membrane protein stability and function.

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Abbreviations

A8-35:

An anionic amphipol of average molecular mass ~4.3 kDa, containing ~35 % free carboxylates, ~25 % octyl side-chains, and ~40 % isopropyl ones

APol:

Amphipol

CAC:

Critical association concentration

DAPol:

A8-35 with per-deuterated side-chains

EINS:

Elastic incoherent neutron scattering

HAPol:

Natural abundance A8-35

INS:

Inelastic neutron scattering

MD:

Molecular dynamics

〈M n〉:

Number-averaged molecular mass

mQ water:

Water purified on a A10 Advantage Millipore System

MSD:

Mean square displacement

OmpA, OmpX:

Respectively outer membrane proteins A and X from Escherichia coli

QENS:

Quasi-elastic neutron scattering

R S :

Stokes radius

SANS:

Small-angle neutron scattering

SEC:

Size exclusion chromatography

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Acknowledgments

Particular thanks are due to Michael Marek Koza and Bernhard Frick, ILL local contacts on IN6 and IN16, respectively. This work was supported by the French Centre National de la Recherche Scientifique (CNRS), by Université Paris–7 Denis Diderot, and by Grant “DYNAMO”, ANR-11-LABX-0011-01 from the French “Initiative d’Excellence” Program. ‘Computational resources were provided by the Minnesota Supercomputing Institute (MSI).

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Authors and Affiliations

  1. Centre for Medical Radiation Physics and Centre for Medical and Molecular Bioscience, University of Wollongong, Wollongong, NSW, 2522, Australia

    Moeava Tehei

  2. Department of Physics, Brandeis University, 415 South Street, Waltham, MA, 02453, USA

    Jason D. Perlmutter

  3. UMR 7099, Centre National de la Recherche Scientifique/Université Paris-7, Institut de Biologie Physico-Chimique (FRC 550), 13 rue Pierre et Marie Curie, 75005, Paris, France

    Fabrice Giusti & Jean-Luc Popot

  4. Department of Biomedical Engineering, University of Minnesota, 312 Church Street SE, Minneapolis, MN, 55455, USA

    Jonathan N. Sachs

  5. Institut de Biologie Structurale, CEA/CNRS/UJF UMR5075, 38027, Grenoble, France

    Giuseppe Zaccai

  6. Institut Laue Langevin, 38042, Grenoble, France

    Giuseppe Zaccai

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  1. Moeava Tehei
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  2. Jason D. Perlmutter
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  3. Fabrice Giusti
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  4. Jonathan N. Sachs
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  5. Giuseppe Zaccai
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  6. Jean-Luc Popot
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Correspondence to Giuseppe Zaccai or Jean-Luc Popot.

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Tehei, M., Perlmutter, J.D., Giusti, F. et al. Thermal Fluctuations in Amphipol A8-35 Particles: A Neutron Scattering and Molecular Dynamics Study. J Membrane Biol 247, 897–908 (2014). https://doi.org/10.1007/s00232-014-9725-1

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