Magnetically Directed Two-Dimensional Crystallization of OmpF Membrane Proteins in Block Copolymers

Steven S. Klara; Patrick O. Saboe; Ian T. Sines; Mahnoush Babaei; Po Lin Chiu; Rita Dezorzi; Kaushik Dayal; Thomas Walz; Manish Kumar; Meagan S. Mauter

doi:10.1021/jacs.5b03320

Magnetically Directed Two-Dimensional Crystallization of OmpF Membrane Proteins in Block Copolymers

Steven S. Klara, Patrick O. Saboe, Ian T. Sines, Mahnoush Babaei, Po Lin Chiu, Rita Dezorzi, Kaushik Dayal, Thomas Walz, Manish Kumar, Meagan S. Mauter

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

26 Scopus citations

Abstract

Two-dimensional (2D) alignment and crystallization of membrane proteins (MPs) is increasingly important in characterizing their three-dimensional (3D) structure, in designing pharmacological agents, and in leveraging MPs for biomimetic devices. Large, highly ordered MP 2D crystals in block copolymer (BCP) matrices are challenging to fabricate, but a facile and scalable technique for aligning and crystallizing MPs in thin-film geometries would rapidly translate into applications. This work introduces a novel method to grow larger and potentially better ordered 2D crystals by performing the crystallization process in the presence of a strong magnetic field. We demonstrate the efficacy of this approach using a β-barrel MP, outer membrane protein F (OmpF), in short-chain polybutadiene-poly(ethylene oxide) (PB-PEO) membranes. Crystals grown in a magnetic field were up to 5 times larger than conventionally grown crystals, and a signal-to-noise (SNR) analysis of diffraction peaks in Fourier transforms of specimens imaged by negative-stain electron microscopy (EM) and cryo-EM showed twice as many high-SNR diffraction peaks, indicating that the magnetic field also improves crystal order.

Original language	English (US)
Pages (from-to)	28-31
Number of pages	4
Journal	Journal of the American Chemical Society
Volume	138
Issue number	1
DOIs	https://doi.org/10.1021/jacs.5b03320
State	Published - Jan 13 2016
Externally published	Yes

ASJC Scopus subject areas

Catalysis
General Chemistry
Biochemistry
Colloid and Surface Chemistry

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title = "Magnetically Directed Two-Dimensional Crystallization of OmpF Membrane Proteins in Block Copolymers",

abstract = "Two-dimensional (2D) alignment and crystallization of membrane proteins (MPs) is increasingly important in characterizing their three-dimensional (3D) structure, in designing pharmacological agents, and in leveraging MPs for biomimetic devices. Large, highly ordered MP 2D crystals in block copolymer (BCP) matrices are challenging to fabricate, but a facile and scalable technique for aligning and crystallizing MPs in thin-film geometries would rapidly translate into applications. This work introduces a novel method to grow larger and potentially better ordered 2D crystals by performing the crystallization process in the presence of a strong magnetic field. We demonstrate the efficacy of this approach using a β-barrel MP, outer membrane protein F (OmpF), in short-chain polybutadiene-poly(ethylene oxide) (PB-PEO) membranes. Crystals grown in a magnetic field were up to 5 times larger than conventionally grown crystals, and a signal-to-noise (SNR) analysis of diffraction peaks in Fourier transforms of specimens imaged by negative-stain electron microscopy (EM) and cryo-EM showed twice as many high-SNR diffraction peaks, indicating that the magnetic field also improves crystal order.",

author = "Klara, {Steven S.} and Saboe, {Patrick O.} and Sines, {Ian T.} and Mahnoush Babaei and Chiu, {Po Lin} and Rita Dezorzi and Kaushik Dayal and Thomas Walz and Manish Kumar and Mauter, {Meagan S.}",

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AU - Klara, Steven S.

AU - Saboe, Patrick O.

AU - Sines, Ian T.

AU - Babaei, Mahnoush

AU - Chiu, Po Lin

AU - Dezorzi, Rita

AU - Dayal, Kaushik

AU - Walz, Thomas

AU - Kumar, Manish

AU - Mauter, Meagan S.

PY - 2016/1/13

Y1 - 2016/1/13

N2 - Two-dimensional (2D) alignment and crystallization of membrane proteins (MPs) is increasingly important in characterizing their three-dimensional (3D) structure, in designing pharmacological agents, and in leveraging MPs for biomimetic devices. Large, highly ordered MP 2D crystals in block copolymer (BCP) matrices are challenging to fabricate, but a facile and scalable technique for aligning and crystallizing MPs in thin-film geometries would rapidly translate into applications. This work introduces a novel method to grow larger and potentially better ordered 2D crystals by performing the crystallization process in the presence of a strong magnetic field. We demonstrate the efficacy of this approach using a β-barrel MP, outer membrane protein F (OmpF), in short-chain polybutadiene-poly(ethylene oxide) (PB-PEO) membranes. Crystals grown in a magnetic field were up to 5 times larger than conventionally grown crystals, and a signal-to-noise (SNR) analysis of diffraction peaks in Fourier transforms of specimens imaged by negative-stain electron microscopy (EM) and cryo-EM showed twice as many high-SNR diffraction peaks, indicating that the magnetic field also improves crystal order.

AB - Two-dimensional (2D) alignment and crystallization of membrane proteins (MPs) is increasingly important in characterizing their three-dimensional (3D) structure, in designing pharmacological agents, and in leveraging MPs for biomimetic devices. Large, highly ordered MP 2D crystals in block copolymer (BCP) matrices are challenging to fabricate, but a facile and scalable technique for aligning and crystallizing MPs in thin-film geometries would rapidly translate into applications. This work introduces a novel method to grow larger and potentially better ordered 2D crystals by performing the crystallization process in the presence of a strong magnetic field. We demonstrate the efficacy of this approach using a β-barrel MP, outer membrane protein F (OmpF), in short-chain polybutadiene-poly(ethylene oxide) (PB-PEO) membranes. Crystals grown in a magnetic field were up to 5 times larger than conventionally grown crystals, and a signal-to-noise (SNR) analysis of diffraction peaks in Fourier transforms of specimens imaged by negative-stain electron microscopy (EM) and cryo-EM showed twice as many high-SNR diffraction peaks, indicating that the magnetic field also improves crystal order.

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Magnetically Directed Two-Dimensional Crystallization of OmpF Membrane Proteins in Block Copolymers

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