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

Mariusz Jaremko, Łukasz Jaremko, Michał Nowakowski, Marek Wojciechowski, Roman H. Szczepanowski, Renata Panecka, Igor Zhukov, Matthias Bochtler, Andrzej Ejchart*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

2 Scopus citations

Abstract

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.

Original languageEnglish (US)
Pages (from-to)69-78
Number of pages10
JournalJournal of Structural Biology
Volume185
Issue number1
DOIs
StatePublished - Jan 2014
Externally publishedYes

Keywords

  • Backbone dynamics
  • First catalytic cysteine half-domain (FCCH)
  • N nuclear magnetic relaxation
  • NMR structure determination
  • NMR study interactions in solution
  • Protein structure
  • Second catalytic cysteine half-domain (SCCH)
  • Ubiquitin-activating enzyme E1

ASJC Scopus subject areas

  • Structural Biology

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