Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2018 Jun 27;13(6):e0199639.
doi: 10.1371/journal.pone.0199639. eCollection 2018.

The crystallization additive hexatungstotellurate promotes the crystallization of the HSP70 nucleotide binding domain into two different crystal forms

Aengus Mac Sweeney  1 , Alain Chambovey  1 , Micha Wicki  1 , Manon Müller  1 , Nadia Artico  1 , Roland Lange  1 , Aleksandar Bijelic  2 , Joscha Breibeck  2 , Annette Rompel  2
Affiliations

The crystallization additive hexatungstotellurate promotes the crystallization of the HSP70 nucleotide binding domain into two different crystal forms

Aengus Mac Sweeney et al. PLoS One. .

Abstract

The use of the tellurium-centered Anderson-Evans polyoxotungstate [TeW6O24]6- (TEW) as a crystallization additive has been described. Here, we present the use of TEW as an additive in the crystallization screening of the nucleotide binding domain (NBD) of HSP70. Crystallization screening of the HSP70 NBD in the absence of TEW using a standard commercial screen resulted in a single crystal form. An identical crystallization screen of the HSP70 NBD in the presence of TEW resulted in both the "TEW free" crystal form and an additional crystal form with a different crystal packing. TEW binding was observed in both crystal forms, either as a well-defined molecule or in overlapping alternate positions suggesting translational disorder. The structures were solved by both molecular replacement and single wavelength anomalous diffraction (SAD) using the anomalous signal of a single bound molecule of TEW. This study adds one more example of TEW binding to a protein and influencing its crystallization behavior.

PubMed Disclaimer

Conflict of interest statement

We have the following interests. Aengus Mac Sweeney, Alain Chambovey, Micha Wicki, Manon Müller, Nadia Artico and Roland Lange are employed by Idorsia Pharmaceuticals Ltd. There are no patents, products in development or marketed products to declare. This does not alter our adherence to all the PLOS ONE policies on sharing data and materials, as detailed online in the guide for authors.

Figures

Fig 1
Fig 1. Chemical structure of TEW.
Fig 2
Fig 2. TEW bound to HSP70 at a crystal contact (crystal form 1).
HSP70 is colored grey, the symmetry related HSP70 is colored yellow, with its bound ADP colored cyan. 2Fo-Fc electron density (contoured at 2 σ) is shown for the TEW molecule.
Fig 3
Fig 3. TEW bound to HSP70 at a crystal contact (crystal form 2, site 1).
HSP70 is colored cyan, the symmetry related HSP70 is colored grey. The two alternate positions are shown with oxygen atoms colored red or yellow. 2Fo-Fc electron density (contoured at 2 σ) is shown for the TEW molecule.
Fig 4
Fig 4. TEW bound to HSP70 at a crystal contact (crystal form 2, site 2).
HSP70 is shown in cyan, the symmetry related HSP70 and its bound ADP are shown in grey. The two alternate positions of TEW are shown with the oxygen atoms colored in red (position A) or yellow (position B). 2Fo-Fc electron density (contoured at 1.5 σ) is shown for the TEW molecule.
Fig 5
Fig 5. Surface potential of crystal form 1 calculated using APBS.
Fig 6
Fig 6. Surface potential of crystal form 2, site 1 calculated using APBS.
Fig 7
Fig 7. Surface potential of crystal form 2, site 2 calculated using APBS.
Fig 8
Fig 8. Thermal unfolding (DSF) curves of HSP70 alone and in the presence of ligands.
HSP70 (Tm = 45.23±0.13 K) and combined with ligands TEW (Tm = 44.84 ± 0.33 K), ADP (Tm = 54.48 ± 0.34 K) or ATP (Tm = 53.3 ± 0.24 K). Lines represent the mean of quadruplicate fluorescence intensity measurements; error bars have been omitted for clarity.
See this image and copyright information in PMC

References

    1. Molitor C, Bijelic A, Rompel A. In situ formation of the first proteinogenically functionalized [TeW6O24O2(Glu)]7- structure reveals unprecedented chemical and geometrical features of the Anderson-type cluster. Chem. Commun. 2016;52: 12286–12289. - PMC - PubMed
    1. Mauracher SG, Molitor C, Al-Oweini R, Kortz U, Rompel A. Crystallization and preliminary X-ray crystallographic analysis of latent isoform PPO4 mushroom (Agaricus bisporus) tyrosinase Acta Crystallogr. F. 2014;70: 263–266. - PMC - PubMed
    1. Mauracher SG, Molitor C, Al-Oweini R, Kortz U, Rompel A. Latent and active abPPO4 mushroom tyrosinases cocrystallized with hexatungstotellurate(VI) in a single crystal. Acta Cryst. D 2014;70: 2301–2315. - PMC - PubMed
    1. Bijelic A, Molitor C, Mauracher SG, Al-Oweini R, Kortz U, Rompel A. Hen egg-white lysozyme crystallisation: protein stacking and structure stability enhanced by a Tellurium(VI)-centred polyoxotungstate. Chembiochem. 2015;16: 233–41. doi: 10.1002/cbic.201402597 - DOI - PMC - PubMed
    1. Bijelic A, Rompel A. Ten Good Reasons for the Use of the Tellurium-Centered Anderson-Evans Polyoxotungstate in Protein Crystallography. Acc. Chem. Res. 2017;50(6): 1441–1448. doi: 10.1021/acs.accounts.7b00109 - DOI - PMC - PubMed

Publication types

Substances