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Comparative Study
. 2016 Oct 1;72(Pt 10):762-771.
doi: 10.1107/S2053230X16014011. Epub 2016 Sep 22.

Comparative analysis of anti-polyglutamine Fab crystals grown on Earth and in microgravity

Gwen E Owens  1 , Danielle M New  1 , Alejandra I Olvera  1 , Julia Ashlyn Manzella  2 , Brittney L Macon  2 , Joshua C Dunn  2 , David A Cooper  2 , Robyn L Rouleau  2 , Daniel S Connor  2 , Pamela J Bjorkman  1
Affiliations
Comparative Study

Comparative analysis of anti-polyglutamine Fab crystals grown on Earth and in microgravity

Gwen E Owens et al. Acta Crystallogr F Struct Biol Commun. .

Abstract

Huntington's disease is one of nine neurodegenerative diseases caused by a polyglutamine (polyQ)-repeat expansion. An anti-polyQ antigen-binding fragment, MW1 Fab, was crystallized both on Earth and on the International Space Station, a microgravity environment where convection is limited. Once the crystals returned to Earth, the number, size and morphology of all crystals were recorded, and X-ray data were collected from representative crystals. The results generally agreed with previous microgravity crystallization studies. On average, microgravity-grown crystals were 20% larger than control crystals grown on Earth, and microgravity-grown crystals had a slightly improved mosaicity (decreased by 0.03°) and diffraction resolution (decreased by 0.2 Å) compared with control crystals grown on Earth. However, the highest resolution and lowest mosaicity crystals were formed on Earth, and the highest-quality crystal overall was formed on Earth after return from microgravity.

Keywords: Huntington's disease; International Space Station; X-ray crystallography; crystallization; huntingtin; microgravity; polyglutamine.

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Figures

Figure 1
Figure 1
Schematics of the HDPCG device used for microgravity crystallization experiments. (a) Handheld HDPCG assembly, (b) HDPCG sample block, (c) HDPCG growth cell in loading configuration, (d) HDPCG growth cell in launch configuration, (e) HDPCG growth cell in microgravity crystal-growth configuration. The scale bar is 5 mm in length.
Figure 2
Figure 2
Timeline of (a) microgravity and (b) ground-control experiments.
Figure 3
Figure 3
Comparison of crystal morphology and size. Each data point represents a one crystal.. Protein crystals grew in both microgravity and ground-control conditions. (a) Morphology of crystals and number of wells containing crystals. Wells 1–5 were set up with MW1 or 3B5H10 Fab, wells 6–10 with Fab + peptide, wells 11 and 12 with HD-16Q, wells 13–22 with Fab + HD-16Q, wells 23–36 with Fab + HD-39Q, wells 37–48 with HD-16Q, HD-25Q, HD-36Q or HD-46Q, wells 49–54 with huntingtin-GFP and wells 55–60 with full-length huntingtin. See Supplementary Table S1 for a complete description of the initial conditions. (b) Area of crystals greater than 400 µm2 grown in microgravity (n = 67) and on Earth (n = 97). Data shown are geometric means with 95% confidence intervals. The geometric mean is suitable for data that range over several orders of magnitude (West et al., 2010 ▸).
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