Abstract��Abstract: The chitin elicitor receptor kinase 1 (CERK1) family of plant immune kinases are pattern recogni-tion receptors (PRRs) capable of sensing bacterial and fungal infections. By recognizing pathogen-associated molecular patterns (PAMPs), such as some pathogen polysaccharides, or symbiotic factors, these receptor pro-teins involve in induction of PAMP-triggered immunity (PTI) or plant-microbe symbiosis. To determine the three-dimensional structure of CERK1 kinase domain, the E. coli-pRSF prokaryotic expression systems of wild-type and mutant AtCERK1 proteins were constructed for overexpression of CERK1 from Arabidopsis. The AtCERK1 proteins with a 6�� His tag were expressed and subsequently purified by nickel column affinity chromatography. Based on the biophysical and biochemical properties of AtCERK1, ion exchange chro-matography and gel filtration chromatography were performed for further purification of the AtCERK1 pro-teins. The resulting AtCERK1 proteins, up to 95% purity, were subjected to screening of crystallization con-ditions. The AtCERK1F460V mutant protein was finally crystallized in the solution containing 0.2 mol/L am-monium fluoride and 20% PEG3350. X-ray diffraction data of these crystals were collected at the highest resolution of 3.2 ?�. In conclusion, the AtCERK1 protein was successfully obtained and crystallized. The preliminary crystallographic analysis of this protein will facilitate future investigation of the structural and functional basis of AtCERK1 mediated plant immunity.
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ţ��Ⱥ, ������, �� ��, ����. ֲ������øAtCERK1����������ᾧ�о�[J]. ������ѧ�о�, 2022, 26(5): 386-395.
NIU Cheng-qun, YANG Zhi-qian, LIN Jie, MING Zhen-hua. Expression and Crystallization of Recombinant Plant Immune Kinase AtCERK1. Life Science Research, 2022, 26(5): 386-395.