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X‐ray crystallographic studies of the extracellular domain of the first plant ATP receptor, DORN1, and the orthologous protein from Camelina sativa
Author(s) -
Li Zhijie,
Chakraborty Sayan,
Xu Guozhou
Publication year - 2016
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 37
ISSN - 2053-230X
DOI - 10.1107/s2053230x16014278
Subject(s) - camelina sativa , extracellular , arabidopsis thaliana , protein kinase a , biochemistry , signal transduction , receptor , biology , microbiology and biotechnology , kinase , chemistry , mutant , gene , ecology , crop
Does not respond to nucleotides 1 (DORN1) has recently been identified as the first membrane‐integral plant ATP receptor, which is required for ATP‐induced calcium response, mitogen‐activated protein kinase activation and defense responses in Arabidopsis thaliana . In order to understand DORN1‐mediated ATP sensing and signal transduction, crystallization and preliminary X‐ray studies were conducted on the extracellular domain of DORN1 (atDORN1‐ECD) and that of an orthologous protein, Camelina sativa lectin receptor kinase I.9 (csLecRK‐I.9‐ECD or csI.9‐ECD). A variety of deglycosylation strategies were employed to optimize the glycosylated recombinant atDORN1‐ECD for crystallization. In addition, the glycosylated csI.9‐ECD protein was crystallized at 291 K. X‐ray diffraction data were collected at 4.6 Å resolution from a single crystal. The crystal belonged to space group C 222 or C 222 1 , with unit‐cell parameters a = 94.7, b = 191.5, c = 302.8 Å. These preliminary studies have laid the foundation for structural determination of the DORN1 and I.9 receptor proteins, which will lead to a better understanding of the perception and function of extracellular ATP in plants.