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Nedd4-2 binding to 14-3-3 modulates the accessibility of its catalytic site and WW domains

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    0556577 - FGÚ 2023 RIV US eng J - Článek v odborném periodiku
    Joshi, Rohit - Pohl, Pavel - Strachotová, D. - Herman, P. - Obšil, Tomáš - Obšilová, Veronika
    Nedd4-2 binding to 14-3-3 modulates the accessibility of its catalytic site and WW domains.
    Biophysical Journal. Roč. 121, č. 7 (2022), s. 1299-1311. ISSN 0006-3495. E-ISSN 1542-0086
    Grant CEP: GA ČR(CZ) GA20-00058S
    Výzkumná infrastruktura: CIISB II - 90127
    Institucionální podpora: RVO:67985823
    Klíčová slova: Nedd4-2 ubiquitin ligase * 14-3-3 protein * fluorescence spectroscopy * protein structure and function
    Obor OECD: Biochemistry and molecular biology
    Impakt faktor: 3.699, rok: 2021

    Neural precursor cells expressed developmentally downregulated protein 4-2 (Nedd4-2), a homologous to the E6-AP carboxyl terminus (HECT) ubiquitin ligase, triggers the endocytosis and degradation of its downstream target molecules by regulating signal transduction through interactions with other targets, including 14-3-3 proteins. In our previous study, we found that 14-3-3 binding induces a structural rearrangement of Nedd4-2 by inhibiting interactions between its structured domains. Here, we used time-resolved fluorescence intensity and anisotropy decay measurements, together with fluorescence quenching and mass spectrometry, to further characterize interactions between Nedd4-2 and 14-3-3 proteins. The results showed that 14-3-3 binding affects the emission properties of AEDANS-labeled WW3, WW4, and, to a lesser extent, WW2 domains, and reduces their mobility, but not those of the WW1 domain, which remains mobile. In contrast, 14-3-3 binding has the opposite effect on the active site of the HECT domain, which is more solvent exposed and mobile in the complexed form than in the apo form of Nedd4-2. Overall, our results suggest that steric hindrance of the WW3 and WW4 domains combined with conformational changes in the catalytic domain may account for the 14-3-3 binding-mediated regulation of Nedd4-2.
    Trvalý link: http://hdl.handle.net/11104/0330738

     
     
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