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

DOI: 10.1371/journal.pone.0221226

Authors: John F. C. Steele (John Innes Centre) , Richard K. Hughes (John Innes Centre) , Mark J. Banfield (John Innes Centre)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Plos One , VOL 14 , PAGES N/A

State: Published (Approved)
Published: August 2019
Diamond Proposal Number(s): 7641

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.

Journal Keywords: Electron density; Protein domains; Gel filtration; Protein structure; Sequence motif analysis; Adenosine triphosphatase; Protein expression; Biophysics

Subject Areas: Biology and Bio-materials, Chemistry


Instruments: I02-Macromolecular Crystallography , I04-Macromolecular Crystallography

Added On: 24/09/2019 11:23

Documents:
bh4hg44fkk4.pdf

Discipline Tags:

Plant science Pathogens Biochemistry Agriculture & Fisheries Chemistry Structural biology Biophysics Life Sciences & Biotech

Technical Tags:

Diffraction Macromolecular Crystallography (MX)