title: X-ray crystallographic studies of the extracellular domain of the first plant ATP receptor, DORN1, and the orthologous protein from Camelina sativa.
abstract: 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 C222 or C2221, 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.
authors.affiliation: Department of Molecular and Structural Biochemistry, North Carolina State University, 26 Polk Hall, Raleigh, NC 27695, USA.
authors.forename: Zhijie
authors.initials: Z
authors.lastname: Li
authors.affiliation: Department of Molecular and Structural Biochemistry, North Carolina State University, 26 Polk Hall, Raleigh, NC 27695, USA.
authors.forename: Sayan
authors.initials: S
authors.lastname: Chakraborty
authors.affiliation: Department of Molecular and Structural Biochemistry, North Carolina State University, 26 Polk Hall, Raleigh, NC 27695, USA.
authors.forename: Guozhou
authors.initials: G
authors.lastname: Xu
mesh_terms: D000255:Adenosine Triphosphate / Q000737:chemistry* / Q000378:metabolism; D000595:Amino Acid Sequence; D017360:Arabidopsis / Q000737:chemistry* / Q000378:metabolism; D029681:Arabidopsis Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D016367:Baculoviridae / Q000235:genetics / Q000378:metabolism; D019607:Brassicaceae / Q000737:chemistry* / Q000378:metabolism; D003001:Cloning, Molecular; D003460:Crystallization; D018360:Crystallography, X-Ray; D015870:Gene Expression; D006031:Glycosylation; D010957:Plasmids / Q000737:chemistry / Q000378:metabolism; D011494:Protein Kinases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D014961:X-Ray Diffraction
chemical_list: D029681:Arabidopsis Proteins; D011994:Recombinant Proteins; D000255:Adenosine Triphosphate; C586088:LecRK-I.9 protein, Arabidopsis; D011494:Protein Kinases
keywords: Arabidopsis thaliana; Camelina sativa; DORN1; glycosylation; lectin receptor kinase I.9; plant ATP receptor