Light-induced structural changes of LOV domain-containing polypeptides from Arabidopsis phototropin 1 and 2 studied by small-angle X-ray scattering

Masayoshi Nakasako, Tatsuya Iwata, Daisuke Matsuoka, Satoru Tokutomi

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Abstract

Phototropin is a blue-light receptor of plants and comprises two light-receptive domains, LOV1 and LOV2, Ser/Thr kinase domain and one linker region connecting the LOV2 and the kinase domains. The LOV2 domain is thought to regulate predominantly the light-dependent autophosphorylation of the kinase domain, leading to cellular signaling cascades. In this study, we constructed recombinant LOV1, LOV2, and LOV2-linker polypeptides from phototropin 1 and phototropin 2 of Arabidopsis thaliana and studied their quaternary structures and light-dependent conformational changes by small-angle X-ray scattering. The molecular weights of the polypeptides determined from scattering intensities demonstrated the dimeric associations of LOV1 polypeptides of both isoforms. In contrast, while LOV2 and LOV2-linker polypeptides of phototropin 1 were homodimers, corresponding polypeptides of phototropin 2 existed as monomeric forms. Under blue-light irradiation, the LOV2-linker polypeptide of phototropin 1 displayed small but definite changes of the scattering profile. Through simulation of low-resolution molecular structures, the changes were likely explained as structural changes of the linker region and/or a movement of the region relative to the LOV2 domain. Light-induced profile changes were not observed in the Cys512Ala mutated LOV2-linker polypeptide of phototropin 1 losing the phototransformation capability. Thus, it was indicated that the photoreaction in the LOV2 domain probably caused the structural changes in the LOV2-linker polypeptide of phototropin 1. On the basis of the results, the interdomain interactions in phototropin are discussed.

Original languageEnglish
Pages (from-to)14881-14890
Number of pages10
JournalBiochemistry
Volume43
Issue number47
DOIs
Publication statusPublished - 2004 Nov 30

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Phototropins
X ray scattering
X-Rays
Light
Peptides
Phosphotransferases
Scattering
Cell signaling
Arabidopsis NPH1 protein
Molecular Structure
Arabidopsis
Molecular structure
Protein Isoforms
Molecular Weight
Molecular weight

ASJC Scopus subject areas

  • Biochemistry

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Light-induced structural changes of LOV domain-containing polypeptides from Arabidopsis phototropin 1 and 2 studied by small-angle X-ray scattering. / Nakasako, Masayoshi; Iwata, Tatsuya; Matsuoka, Daisuke; Tokutomi, Satoru.

In: Biochemistry, Vol. 43, No. 47, 30.11.2004, p. 14881-14890.

Research output: Contribution to journalArticle

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