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NMR structural studies of the first catalytic half-domain of ubiquitin activating enzyme

Jaremko, Mariusz and Jaremko, Łukasz and Nowakowski, Michał and Wojciechowski, Marek and Szczepanowski, Roman H. and Panecka, Renata and Zhukov, Igor and Bochtler, Matthias and Ejchart, Andrzej (2014) NMR structural studies of the first catalytic half-domain of ubiquitin activating enzyme. Journal of Structural Biology, 185 (1). pp. 69-78. ISSN 1047-8477

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Official URL: http://www.journals.elsevier.com/journal-of-struct...


We report a high resolution NMR structure and 15N relaxation studies of the first catalytic cysteine half-domain (FCCH) of the mouse ubiquitin-activating enzyme E1, together with interaction studies of FCCH and the other catalytic E1 subdomain - SCCH (second catalytic cysteine half-domain). In solution, mouse FCCH forms a well-defined six-stranded antiparallel β-barrel structure, a common fold for many proteins with a variety of cellular functions. 15N relaxation data reveal FCCH complex backbone dynamics and indicate which residues experience slow intramolecular motions. Some of these residues make contacts with the polar face of ubiquitin in the co-crystal structure of yeast E1 and ubiquitin. However, the titration of FCCH with ubiquitin does not show any visible chemical shift changes in the 2D 1H/15N HSQC spectra of the FCCH. The 2D 1H/15N HSQC experiments performed both for each catalytic half-domain individually and for their equimolar mixture in the milimolar concentration range display no detectable chemical shift perturbation, suggesting a lack of interaction between the two subdomains unless they are covalently linked via the adenylation domain.

Item Type:Article
Subjects:Q Science > QH Natural history > QH301 Biology
Divisions:NMR Laboratory
ID Code:676
Deposited By: professor Andrzej Ejchart
Deposited On:16 Apr 2014 08:25
Last Modified:21 Oct 2016 09:23

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