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

Mariusz Jaremko, Lukasz 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 15 N 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. 15 N 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 1 H/ 15 N HSQC spectra of the FCCH. The 2D 1 H/ 15 N 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 1 2014

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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