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Oil-free hyaluronic acid matrix for serial femtosecond crystallography.

Sugahara M, Song C, Suzuki M, Masuda T, Inoue S, Nakane T, Yumoto F, Nango E, Tanaka R, Tono K, Joti Y, Kameshima T, Hatsui T, Yabashi M, Nureki O, Numata K, Iwata S - Sci Rep (2016)

Bottom Line: The grease matrix was originally introduced as a microcrystal-carrier for serial femtosecond crystallography and has been expanded to applications for various types of proteins, including membrane proteins.However, the grease-based matrix has limited application for oil-sensitive proteins.Here we introduce a grease-free, water-based hyaluronic acid matrix.

View Article: PubMed Central - PubMed

Affiliation: RIKEN SPring-8 Center, Kouto, Sayo-cho, Sayo-gun, Hyogo 679-5148, Japan.

ABSTRACT
The grease matrix was originally introduced as a microcrystal-carrier for serial femtosecond crystallography and has been expanded to applications for various types of proteins, including membrane proteins. However, the grease-based matrix has limited application for oil-sensitive proteins. Here we introduce a grease-free, water-based hyaluronic acid matrix. Applications for proteinase K and lysozyme proteins were able to produce electron density maps at 2.3-Å resolution.

No MeSH data available.


Related in: MedlinePlus

Typical XFEL single diffraction patterns from three carriers.(a) Mineral oil-based AZ grease, (b) Super Lube synthetic grease, and (c) hyaluronic acid. Resolution at the edges corresponds to ~2.3 Å (dashed circle). (d) The average background scattering intensities of ~2,000 images from each matrix. AZ grease, Super Lube synthetic grease and hyaluronic acid are depicted by the black, blue and green lines, respectively.
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f1: Typical XFEL single diffraction patterns from three carriers.(a) Mineral oil-based AZ grease, (b) Super Lube synthetic grease, and (c) hyaluronic acid. Resolution at the edges corresponds to ~2.3 Å (dashed circle). (d) The average background scattering intensities of ~2,000 images from each matrix. AZ grease, Super Lube synthetic grease and hyaluronic acid are depicted by the black, blue and green lines, respectively.

Mentions: We carried out SFX experiments using femtosecond X-ray pulses from the SPring-8 Angstrom Compact Free Electron Laser (SACLA)6. The X-ray wavelength was kept at 1.77 Å (7 keV). At first, the background scatterings from three crystal matrix carriers, a mineral oil-based AZ grease13, a synthetic grease Super Lube1718 and a hyaluronic acid aqueous solution, were compared. Grease generated diffuse scatterings in the resolution range of 4–5 Å (Fig. 1a,b). AZ grease gave a diffraction ring at ~14-Å resolution. For the Super Lube grease, we observed a diffraction ring at ~4.8-Å resolution in about 30% of all diffraction images (Supplementary Fig. 1). Weaker background scattering was noted when using hyaluronic acid compared with those of greases (Fig. 1c,d). In this study, using two carriers, synthetic grease Super Lube and hyaluronic acid matrices, we investigated proteinase K (5–10 μm) and lysozyme (7–10 μm) crystals (Supplementary Fig. 2) to demonstrate their general applicability as crystal carriers. A flow rate of 0.48 μl/min was used for all samples. The grease and hyaluronic acid matrices formed a stable flow for all protein samples (Supplementary Fig. 3).


Oil-free hyaluronic acid matrix for serial femtosecond crystallography.

Sugahara M, Song C, Suzuki M, Masuda T, Inoue S, Nakane T, Yumoto F, Nango E, Tanaka R, Tono K, Joti Y, Kameshima T, Hatsui T, Yabashi M, Nureki O, Numata K, Iwata S - Sci Rep (2016)

Typical XFEL single diffraction patterns from three carriers.(a) Mineral oil-based AZ grease, (b) Super Lube synthetic grease, and (c) hyaluronic acid. Resolution at the edges corresponds to ~2.3 Å (dashed circle). (d) The average background scattering intensities of ~2,000 images from each matrix. AZ grease, Super Lube synthetic grease and hyaluronic acid are depicted by the black, blue and green lines, respectively.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4834484&req=5

f1: Typical XFEL single diffraction patterns from three carriers.(a) Mineral oil-based AZ grease, (b) Super Lube synthetic grease, and (c) hyaluronic acid. Resolution at the edges corresponds to ~2.3 Å (dashed circle). (d) The average background scattering intensities of ~2,000 images from each matrix. AZ grease, Super Lube synthetic grease and hyaluronic acid are depicted by the black, blue and green lines, respectively.
Mentions: We carried out SFX experiments using femtosecond X-ray pulses from the SPring-8 Angstrom Compact Free Electron Laser (SACLA)6. The X-ray wavelength was kept at 1.77 Å (7 keV). At first, the background scatterings from three crystal matrix carriers, a mineral oil-based AZ grease13, a synthetic grease Super Lube1718 and a hyaluronic acid aqueous solution, were compared. Grease generated diffuse scatterings in the resolution range of 4–5 Å (Fig. 1a,b). AZ grease gave a diffraction ring at ~14-Å resolution. For the Super Lube grease, we observed a diffraction ring at ~4.8-Å resolution in about 30% of all diffraction images (Supplementary Fig. 1). Weaker background scattering was noted when using hyaluronic acid compared with those of greases (Fig. 1c,d). In this study, using two carriers, synthetic grease Super Lube and hyaluronic acid matrices, we investigated proteinase K (5–10 μm) and lysozyme (7–10 μm) crystals (Supplementary Fig. 2) to demonstrate their general applicability as crystal carriers. A flow rate of 0.48 μl/min was used for all samples. The grease and hyaluronic acid matrices formed a stable flow for all protein samples (Supplementary Fig. 3).

Bottom Line: The grease matrix was originally introduced as a microcrystal-carrier for serial femtosecond crystallography and has been expanded to applications for various types of proteins, including membrane proteins.However, the grease-based matrix has limited application for oil-sensitive proteins.Here we introduce a grease-free, water-based hyaluronic acid matrix.

View Article: PubMed Central - PubMed

Affiliation: RIKEN SPring-8 Center, Kouto, Sayo-cho, Sayo-gun, Hyogo 679-5148, Japan.

ABSTRACT
The grease matrix was originally introduced as a microcrystal-carrier for serial femtosecond crystallography and has been expanded to applications for various types of proteins, including membrane proteins. However, the grease-based matrix has limited application for oil-sensitive proteins. Here we introduce a grease-free, water-based hyaluronic acid matrix. Applications for proteinase K and lysozyme proteins were able to produce electron density maps at 2.3-Å resolution.

No MeSH data available.


Related in: MedlinePlus