Interdomain Flexibility within NADPH Oxidase Suggested by SANS Using LMNG Stealth Carrier - Groupe Membrane et pathogènes / Membrane and Pathogens Group (IBS-MP) Accéder directement au contenu
Article Dans Une Revue Biophysical Journal Année : 2020

Interdomain Flexibility within NADPH Oxidase Suggested by SANS Using LMNG Stealth Carrier

Résumé

Small angle neutron scattering (SANS) provides a method to obtain important low-resolution information for integral membrane proteins (IMPs), challenging targets for structural determination. Specific deuteration furnishes a "stealth" carrier for the solubilized IMP. We used SANS to determine a structural envelope of SpNOX, the Streptococcus pneumoniae NADPH oxidase (NOX), a prokaryotic model system for exploring structure and function of eukaryotic NOXes. SpNOX was solubilized in the detergent lauryl maltose neopentyl glycol, which provides optimal SpNOX stability and activity. Using deuterated solvent and protein, the lauryl maltose neopentyl glycol was experimentally undetected in SANS. This affords a cost-effective SANS approach for obtaining novel structural information on IMPs. Combining SANS data with molecular modeling provided a first, to our knowledge, structural characterization of an entire NOX enzyme. It revealed a distinctly less compact structure than that predicted from the docking of homologous crystal structures of the separate transmembrane and dehydrogenase domains, consistent with a flexible linker connecting the two domains.
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Dates et versions

hal-02921448 , version 1 (18-11-2020)

Identifiants

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Annelise Vermot, Isabelle Petit-Härtlein, Cécile Breyton, Aline Le Roy, Michel Thépaut, et al.. Interdomain Flexibility within NADPH Oxidase Suggested by SANS Using LMNG Stealth Carrier. Biophysical Journal, 2020, 119 (3), pp.605-618. ⟨10.1016/j.bpj.2020.06.025⟩. ⟨hal-02921448⟩
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