TY - JOUR
T1 - 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
AU - Alexandre, M.T.A.
AU - van Grondelle, R.
AU - Hellingwerf, K.J.
AU - Kennis, J.T.M.
N1 - 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
PY - 2009
Y1 - 2009
N2 - 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
AB - 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
U2 - 10.1016/j.bpj.2009.03.047
DO - 10.1016/j.bpj.2009.03.047
M3 - Article
SN - 0006-3495
VL - 97
SP - 238
EP - 247
JO - Biophysical Journal
JF - Biophysical Journal
IS - 1
ER -