An integrative protocol for the structure determination of the mouse ASC-PYD filament

Mazur, Adam and Broz, Petr and Hiller, Sebastian. (2019) An integrative protocol for the structure determination of the mouse ASC-PYD filament. In: Methods in Enzymology, 625. New York, pp. 205-222.

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Official URL: https://edoc.unibas.ch/75745/

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Several inflammasomes that activate as part of the eukaryotic innate immune response contain long helical filaments of the adaptor protein ASC as a central structural element. Here, we describe a technical protocol that has enabled the structure determination of the filament of the ASC pyrin domain. The protocol integrates data from cryo-electron microscopy and solid-state NMR spectroscopy into a single simulated annealing protocol to determine structural coordinates that fit all input data optimally. The structure shows that the ASC pyrin domain filament is formed by helical stacking of individual pyrin domains forms and that the CARD domains are flexibly attached to the filament outside. An artificial perturbation of the input data shows that the integrated structure determination protocol can allow high quality structures even at resolutions of the electron density map as low 8 Å. The protocol is extendable to other structural input data from biochemical or biophysical experiments.
Faculties and Departments:05 Faculty of Science > Departement Biozentrum > Structural Biology & Biophysics > Structural Biology (Hiller)
05 Faculty of Science > Departement Biozentrum > Services Biozentrum > Research IT (Podvinec)
UniBasel Contributors:Hiller, Sebastian and Podvinec, Michael and Mazur, Adam
Item Type:Book Section
Book Section Subtype:Further Contribution in a Book
Note:Publication type according to Uni Basel Research Database: Book item
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Last Modified:22 Jan 2022 04:12
Deposited On:24 Mar 2020 10:52

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