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Elucidating the Signal Transduction Mechanism of the Blue-Light-Regulated Photoreceptor YtvA: From Photoactivation to Downstream Regulation

ACS Chemical Biology. 2024-02; 
YongLe He, Jinnette Tolentino Collado, James N Iuliano, Helena A Woroniecka, Christopher R Hall, Agnieszka A Gil, Sergey P Laptenok, Gregory M Greetham, Boris Illarionov, Adelbert Bacher, Markus Fischer, Jarrod B French, Andras Lukacs, Stephen R Meech, Peter J Tonge
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Gene Synthesis … cloned into a pET15b vector (Novagen) in the frame with an N-terminal six-His tag. The gene encoding B. japonicum FixJ was synthesized and cloned into a pET28c vector (GenScript). … Get A Quote

摘要

The blue-light photoreceptor YtvA from has an N-terminal flavin mononucleotide (FMN)-binding light-oxygen-voltage (LOV) domain that is fused to a C-terminal sulfate transporter and anti-σ factor antagonist (STAS) output domain. To interrogate the signal transduction pathway that leads to photoactivation, the STAS domain was replaced with a histidine kinase, so that photoexcitation of the flavin could be directly correlated with biological activity. N94, a conserved Asn that is hydrogen bonded to the FMN C2═O group, was replaced with Ala, Asp, and Ser residues to explore the role of this residue in triggering the structural dynamics that activate the output domain. Femtosecond to millisecond time-resolved mu... More

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