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Tryptophan Fluorescence in the Bacillus subtilis Phototropin‐related Protein YtvA as a Marker of Interdomain Interaction ¶
Author(s) -
Losi Aba,
Ternelli Elena,
Gärtner Wolfgang
Publication year - 2004
Publication title -
photochemistry and photobiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.818
H-Index - 131
eISSN - 1751-1097
pISSN - 0031-8655
DOI - 10.1111/j.1751-1097.2004.tb00063.x
Subject(s) - phototropin , flavin mononucleotide , fluorescence , chemistry , tryptophan , förster resonance energy transfer , photochemistry , bacillus subtilis , covalent bond , flavin group , mutant , amino acid , biology , biochemistry , physics , arabidopsis thaliana , genetics , organic chemistry , quantum mechanics , bacteria , gene , enzyme
ABSTRACT The Bacillus subtilis protein YtvA, related to plant phototropins (phot), binds flavin mononucleotide (FMN) within the N‐terminal light, oxygen and voltage (LOV) domain. The blue light‐triggered photocycle of YtvA and phot involves the reversible formation of a covalent photoadduct between FMN and a cysteine (cys) residue. YtvA contains a single tryptophan, W103, localized on the LOV domain and conserved in all phot‐LOV domains. In this study, we show that the fluorescence parameters of W103 in YtvA‐LOV are markedly different from those observed in the full‐length YtvA. The fluorescence quantum yields are ca 0.03 and 0.08, respectively. In YtvA‐LOV, the maximum is redshifted ( ca 345 vs 335 nm) and the average fluorescence lifetime shorter (2.7 vs 4.7 ns). These data indicate that W103 is located in a site of tight contact between the two domains of YtvA. In the FMN‐cys adduct, selective excitation of W103 at 295 nm results in minimal changes of the fluorescence parameters with respect to the dark state. On 280 nm excitation, however, there is a detectable decrease in the fluorescence emitted from tyrosines, with concomitant increase in W103 fluorescence. This effect is reversible in the dark and might arise from a light‐regulated energy transfer process from a yet unidentified tyrosine to W103.