AtNPR4 from Arabidopsis thaliana: expression, purification, crystallization and crystallographic analysis

作者：Yang, Qingzhan; Zhang, Mengze; Zheng, Jimin^*; Jia, Zongchao^*

来源：Acta Crystallographica Section F-Structural Biology Communications, 2019, 75(Pt 1): 67-72.

DOI：10.1107/S2053230X18017235

• salicylic acid
• systemic acquired resistance
• protein purification
• regulatory mechanism
• crystal diffraction
• Arabidopsis thaliana

Salicylic acid (SA) is an important phytohormone that is involved in the regulation of plant defence, growth and development. A large number of proteins have been shown to have the ability to interact with SA, and NPR4 has been demonstrated to be a receptor of SA that plays significant roles in the innate immune response of plants. In this study, Spodoptera frugiperda (Sf9) cells were used to express full-length AtNPR4 from Arabidopsis thaliana. To facilitate crystallization, T4 lysozyme (T4L) was added to the N-terminus of the AtNPR4 protein. The recombinant T4L-AtNPR4 protein was expressed, purified and crystallized using the sitting-drop and hanging-drop vapour-diffusion methods. The T4L-AtNPR4 crystals have symmetry consistent with space group C2, with unit-cell parameters a = 93.7, b = 85.8, c = 88.2 angstrom, beta = 90 degrees and one molecule per asymmetric unit. The best crystal diffracted to a resolution of 2.75 angstrom. Structure determination is in progress.

• 出版日期2019-1
• 单位北京师范大学