Conformational Heterogeneity and Propagation of Structural Changes in the LOV2/J alpha Domain from Avena sativa Phototropin 1 as Recorded by Temperature-Dependent FTIR Spectroscopy

M.T.A. Alexandre, R. van Grondelle, K.J. Hellingwerf, J.T.M. Kennis

Research output: Contribution to JournalArticleAcademicpeer-review

Abstract

Phototropins control phototropism, chloroplast movement, stomatal opening, and leaf expansion in plants. Phototropin 1 (phot1) is composed of a kinase domain linked to two blue light-sensing domains, LOV2 and LOV1, which bind flavin mononucleotide. Disruption of the interaction between the LOV2 domain and a helical segment named Jα, joining LOV to the kinase domain, induces the subsequent kinase activity of phototropin 1 and further-downstream signal transduction. Here we study the effects of temperature and hydration on the light-triggered signal propagation in the phot1 LOV2 domain of Avena sativa (AsLOV2/Jα), using Fourier transform infrared spectroscopy to unravel part of the molecular mechanism of phototropin 1. We report that AsLOV2/Ja shows an intense signal in the amide I and II regions, arising mainly from β-sheet changes and the unbinding of the Jα helix from the Per-ARNT-Sim core and its subsequent partial unfolding. Importantly, these structural changes only occur under conditions of full hydration and at temperatures above 280 K. We characterized a newly isolated low-hydration intermediate that shows a downshift of high-frequency amide I signals and that possibly corresponds to loop tightening, without large β-sheet or Ja structural changes. In addition, we report a heterogeneity in AsLOV2/Jα involving two different C(4)=O conformer populations, coexisting in the dark state and characterized by C(4)=Ocarbonyl frequencies at 1712 cm
Original languageEnglish
Pages (from-to)238-247
JournalBiophysical Journal
Volume97
Issue number1
DOIs
Publication statusPublished - 2009

Bibliographical note

Conformational Heterogeneity and Propagation of Structural Changes in the LOV2/J alpha Domain from Avena sativa Phototropin 1 as Recorded by Temperature-Dependent FTIR Spectroscopy

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