期刊
PHOTOCHEMICAL & PHOTOBIOLOGICAL SCIENCES
卷 21, 期 9, 页码 1627-1636出版社
SPRINGERNATURE
DOI: 10.1007/s43630-022-00245-9
关键词
Knotless phytochrome; Photochemistry; Time-resolved spectroscopy; Flash photolysis
资金
- Deutsche Forschungsgemeinschaft [WA 1850/4-3, 417685888]
This study compares the photodynamics of knotless phytochromes with and without the PHY domain, revealing that the presence of the PHY domain slows down early dynamics and binding pocket relaxation. The photoproduct formation is also affected by the PHY domain and shows pH dependence.
The ability of some knotless phytochromes to photoconvert without the PHY domain allows evaluation of the distinct effect of the PHY domain on their photodynamics. Here, we compare the ms dynamics of the single GAF domain (g1) and the GAF-PHY (g1g2) construct of the knotless phytochrome A112699 from cyanobacterium Nostoc punctiforme. While the spectral signatures and occurrence of the intermediates are mostly unchanged by the domain composition, the presence of the PHY domain slows down the early forward and reverse dynamics involving chromophore and protein binding pocket relaxation. We assign this effect to a more restricted binding pocket imprinted by the PHY domain. The photoproduct formation is also slowed down by the presence of the PHY domain but to a lesser extent than the early dynamics. This indicates a rate limiting step within the GAF and not the PHY domain. We further identify a pH dependence of the biphasic photoproduct formation hinting towards a pKa dependent tuning mechanism. Our findings add to the understanding of the role of the individual domains in the photocycle dynamics and provide a basis for engineering of phytochromes towards biotechnological applications. [GRAPHICS] .
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