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.
Magnetically Directed Two-Dimensional Crystallization of OmpF Membrane Proteins in Block Copolymers
DE ZORZI, RITA;
2016-01-01
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.File | Dimensione | Formato | |
---|---|---|---|
jacs.5b03320.pdf
Accesso chiuso
Tipologia:
Documento in Versione Editoriale
Licenza:
Digital Rights Management non definito
Dimensione
2.14 MB
Formato
Adobe PDF
|
2.14 MB | Adobe PDF | Visualizza/Apri Richiedi una copia |
ja5b03320_si_001.pdf
Accesso chiuso
Descrizione: Supporting information
Tipologia:
Altro materiale allegato
Licenza:
Digital Rights Management non definito
Dimensione
4.62 MB
Formato
Adobe PDF
|
4.62 MB | Adobe PDF | Visualizza/Apri Richiedi una copia |
2872210_ja5b03320_si_001-PostPrint.pdf
accesso aperto
Descrizione: Post Print VQR3
Tipologia:
Bozza finale post-referaggio (post-print)
Licenza:
Digital Rights Management non definito
Dimensione
4.59 MB
Formato
Adobe PDF
|
4.59 MB | Adobe PDF | Visualizza/Apri |
2872210_jacs.5b03320-PostPrint.pdf
accesso aperto
Descrizione: Post Print VQR3
Tipologia:
Bozza finale post-referaggio (post-print)
Licenza:
Digital Rights Management non definito
Dimensione
2.66 MB
Formato
Adobe PDF
|
2.66 MB | Adobe PDF | Visualizza/Apri |
Pubblicazioni consigliate
I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.