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Phototropism pp 175-190 | Cite as

Measurements of Photoreaction and Kinase Activity of Phototropin, a Photoreceptor Protein for Tropic Response in Plants: Involvement of Kinase Activity in the Photosensitivity of Tropic Response

  • Koji Okajima
  • Satoru TokutomiEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1924)

Abstract

Phototropin is a photoreceptor protein responsible for phototropic responses in plants. A phototropin molecule has two photoreceptive domains named LOV1 and LOV2 in the N-terminal region. Blue light absorbed by a chromophore in these domains triggers conformational changes in the protein moiety. The C-terminal region of phototropin forms a Ser/Thr kinase that is activated by these conformational changes. The activated phototropin kinase transmits signals downstream leading to tropic responses. The lifetime of the activated state may concern the sensitivity of the tropic responses to light. Thus, spectrophotometric and kinase activity analyses of phototropin are important to understand the light signaling processes related to the photosensitivity. The preparation of polypeptide samples of Arabidopsis phototropin and the methods of spectroscopic measurements and kinase assay of these samples are shown in this chapter.

Key words

Kinase assay LOV domain Photoreaction Phototropin Ser/Thr kinase Spectrophotometry 

Notes

Acknowledgments

This work was supported in part by Grant-in-Aid for Scientific Research on Innovative Areas to S.T. (22120005) and Grant-in-Aid for Scientific Research on Priority Areas to S.T. (17084008) from the Ministry of Education, Culture, Sports, Science, and Technology, Japan, and Grant-in-Aid for Exploratory Research to S.T. (23657105) and Grant-in-aid for Young Scientist B to K.O. (15K18559) from the Japan Society for the Promotion of Science.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of Physics, Faculty of Science and TechnologyKeio UniversityHiyoshiJapan
  2. 2.Department of Biological Science, Graduate School of ScienceOsaka Prefecture UniversitySakaiJapan
  3. 3.Botanical GardensTohoku UniversitySendaiJapan

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