Structural and biochemical studies of an NB-ARC domain from a plant NLR immune receptor

PLoS One. 2019 Aug 28;14(8):e0221226. doi: 10.1371/journal.pone.0221226. eCollection 2019.

Abstract

Plant NLRs are modular immune receptors that trigger rapid cell death in response to attempted infection by pathogens. A highly conserved nucleotide-binding domain shared with APAF-1, various R-proteins and CED-4 (NB-ARC domain) is proposed to act as a molecular switch, cycling between ADP (repressed) and ATP (active) bound forms. Studies of plant NLR NB-ARC domains have revealed functional similarities to mammalian homologues, and provided insight into potential mechanisms of regulation. However, further advances have been limited by difficulties in obtaining sufficient yields of protein suitable for structural and biochemical techniques. From protein expression screens in Escherichia coli and Sf9 insect cells, we defined suitable conditions to produce the NB-ARC domain from the tomato NLR NRC1. Biophysical analyses of this domain showed it is a folded, soluble protein. Structural studies revealed the NRC1 NB-ARC domain had co-purified with ADP, and confirmed predicted structural similarities between plant NLR NB-ARC domains and their mammalian homologues.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine Triphosphatases / chemistry
  • Adenosine Triphosphatases / genetics
  • Amino Acid Sequence / genetics
  • Animals
  • Chromatography, Gel
  • Disease Resistance / genetics
  • Escherichia coli / genetics
  • Gene Expression Regulation, Plant
  • Insecta / cytology
  • Models, Molecular
  • Nucleotide Motifs / genetics
  • Plant Diseases / genetics*
  • Plant Immunity / genetics*
  • Plant Proteins / chemistry
  • Plant Proteins / genetics*
  • Protein Domains / genetics
  • Protein Folding
  • Solanum lycopersicum / chemistry*
  • Solanum lycopersicum / genetics

Substances

  • Plant Proteins
  • Adenosine Triphosphatases