Alpha proton detection based backbone assignment of intrinsically disordered proteins

Methods Mol Biol. 2012:895:211-26. doi: 10.1007/978-1-61779-927-3_15.

Abstract

Assignment of NMR resonance frequencies to a particular atom in the molecule establishes a vital step for any detailed structural study. Approaches for sequential assignment typically involve amide proton detection, which may become suboptimal in case of intrinsically disordered proteins (IDPs) at high pH and/or temperature. Here we describe an alternative approach: assignment protocol based on alpha proton detected triple-resonance experiments, which offer several advantages over well-established experiments relying on amide proton detection. Our experiments are suitable for studies of IDPs at any pH and enable sequential assignment of proline-rich segments.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acids / chemistry
  • Bacterial Proteins / chemistry
  • Humans
  • Melanoma-Specific Antigens / chemistry*
  • Nuclear Magnetic Resonance, Biomolecular*
  • Protein Conformation
  • Protons
  • Quantum Theory

Substances

  • Amino Acids
  • Bacterial Proteins
  • IgG Fc-binding protein, Streptococcus
  • Melanoma-Specific Antigens
  • Protons
  • melanoma-derived CT16 cancer testis antigen, human