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Using HEK293T Expression System to Study Photoactive Plant Cryptochromes.

  • Author(s): Yang, Liang
  • Wang, Xu
  • Deng, Weixian
  • Mo, Weiliang
  • Gao, Jie
  • Liu, Qing
  • Zhang, Chuanyu
  • Wang, Qin
  • Lin, Chentao
  • Zuo, Zecheng
  • et al.
Abstract

Cryptochromes are photolyase-like blue light receptors that are conserved in plants and animals. Although the light-dependent catalytic mechanism of photolyase is well studied, the photochemical mechanism of cryptochromes remains largely unknown. Lack of an appropriate protein expression system to obtain photochemically active cryptochrome holoproteins is a technical obstacle for the study of plant cryptochromes. We report here an easy-to-use method to express and study Arabidopsis cryptochrome in HEK293T cells. Our results indicate that Arabidopsis cryptochromes expressed in HEK293T are photochemically active. We envision a broad use of this method in the functional investigation of plant proteins, especially in the large-scale analyses of photochemical activities of cryptochromes such as blue light-dependent protein-protein interactions.

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