1H, 13C and 15N assignments of Sgt2 N-terminal dimerisation domain and its binding partner, Get5 Ubiquitin-like domain

Biomol NMR Assign. 2013 Oct;7(2):271-4. doi: 10.1007/s12104-012-9425-7. Epub 2012 Sep 22.

Abstract

The first stage of the GET (guided entry of tail-anchored proteins) mechanism for tail-anchored (TA) membrane protein insertion is thought to occur when Sgt2 (small, glutamine-rich, tetratricopeptide repeat-containing protein 2) binds TA proteins upon their release from the ribosome. It sorts them and passes the majority over to a complex of Get5 and Get4 for transmission along the GET pathway and delivery to their membrane destination. Sgt2 is a 38 kDa protein consisting of three domains. The N-terminal domain effects tight dimerisation of the protein and is also the site for binding with the ubiquitin-like (UBL) domain of Get5. Here we have expressed and purified uniformly-(15)N/(13)C-labelled N-terminal Sgt2 (Sgt2_NT) and its binding partner, Get5 UBL domain (Get5_UBL) and assigned the backbone and side-chain resonances as a basis for structure solution of the individual components and, ultimately, the complex. This will provide detailed molecular insight into the early stages of the GET pathway.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Carbon Isotopes
  • Carrier Proteins / chemistry*
  • Carrier Proteins / metabolism*
  • Molecular Sequence Data
  • Nitrogen Isotopes
  • Nuclear Magnetic Resonance, Biomolecular*
  • Protein Binding
  • Protein Multimerization*
  • Protein Structure, Tertiary
  • Protons*
  • Saccharomyces cerevisiae / metabolism
  • Saccharomyces cerevisiae Proteins / chemistry*
  • Saccharomyces cerevisiae Proteins / metabolism*
  • Ubiquitin / chemistry*
  • Ubiquitin / metabolism*

Substances

  • Carbon Isotopes
  • Carrier Proteins
  • Mdy2 protein, S cerevisiae
  • Nitrogen Isotopes
  • Protons
  • Saccharomyces cerevisiae Proteins
  • Sgt2 protein, S cerevisiae
  • Ubiquitin