Preliminary crystallographic analysis of the ankyrin-repeat domain of Arabidopsis thaliana AKT1: identification of the domain boundaries for protein crystallization.
The Arabidopsis thaliana K(+) transporter 1 (AKT1) participates in the maintenance of an adequate cell potassium (K(+)) concentration. The CBL-interacting protein kinase 23 (CIPK23) activates AKT1 for K(+) uptake under low-K(+) conditions. This process is mediated by the interaction between the cytosolic ankyrin-repeat (AR) domain of AKT1 and the kinase domain of CIPK23. However, the precise boundaries of the AR domain and the residues responsible for the interaction are still unknown. Here, the optimization procedure to obtain an AR domain construct suitable for crystallization and the preliminary crystallographic analysis of the obtained crystals are reported. The crystals belonged to space group P21212, with unit-cell parameters a = 34.83, b = 65.89, c = 85.44 Å, and diffracted to 1.98 Å resolution.
Departamento de Cristalografía y Biología Estructural, Instituto de Química-Física `Rocasolano', CSIC, Serrano 119, 28006 Madrid, Spain.
Antonio
A
Chaves-Sanjuán
Departamento de Cristalografía y Biología Estructural, Instituto de Química-Física `Rocasolano', CSIC, Serrano 119, 28006 Madrid, Spain.
María José
MJ
Sánchez-Barrena
Departamento de Cristalografía y Biología Estructural, Instituto de Química-Física `Rocasolano', CSIC, Serrano 119, 28006 Madrid, Spain.
Juana María
JM
González-Rubio
Departamento de Cristalografía y Biología Estructural, Instituto de Química-Física `Rocasolano', CSIC, Serrano 119, 28006 Madrid, Spain.
Armando
A
Albert
D017089:Ankyrin Repeat; D017360:Arabidopsis / Q000378:metabolism*; D029681:Arabidopsis Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D003460:Crystallization; D018360:Crystallography, X-Ray / Q000379:methods*; D015221:Potassium Channels / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
D029681:Arabidopsis Proteins; D015221:Potassium Channels; C074500:AKT1 protein, Arabidopsis
AKT1; Arabidopsis thaliana; K+ transporter 1