Tuning flavin environment to detect and control light-induced conformational switching in Drosophila cryptochrome
| العنوان: | Tuning flavin environment to detect and control light-induced conformational switching in Drosophila cryptochrome |
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| المؤلفون: | Brian R. Crane, Connor M Schneps, Siddarth Chandrasekaran, Changfan Lin, Cristina C DeOliveira, Abir Ganguly, Robert Dunleavy |
| المصدر: | Communications Biology, Vol 4, Iss 1, Pp 1-12 (2021) Communications Biology |
| بيانات النشر: | Springer Science and Business Media LLC, 2021. |
| سنة النشر: | 2021 |
| مصطلحات موضوعية: | Protein Denaturation, Nitroxide mediated radical polymerization, Light, Semiquinone, Protein Conformation, QH301-705.5, Timeless, Medicine (miscellaneous), Flavin group, Plasma protein binding, Molecular Dynamics Simulation, Article, General Biochemistry, Genetics and Molecular Biology, Structure-Activity Relationship, 03 medical and health sciences, 0302 clinical medicine, Protein structure, Cryptochrome, Flavins, Benzoquinones, Animals, Drosophila Proteins, Biology (General), Eye Proteins, 030304 developmental biology, 0303 health sciences, Chemistry, Electron Spin Resonance Spectroscopy, Circadian rhythm signalling peptides and proteins, Molecular conformation, Cryptochromes, Drosophila melanogaster, Sortase A, Biophysics, General Agricultural and Biological Sciences, 030217 neurology & neurosurgery, Protein Binding |
| الوصف: | Light-induction of an anionic semiquinone (SQ) flavin radical in Drosophila cryptochrome (dCRY) alters the dCRY conformation to promote binding and degradation of the circadian clock protein Timeless (TIM). Specific peptide ligation with sortase A attaches a nitroxide spin-probe to the dCRY C-terminal tail (CTT) while avoiding deleterious side reactions. Pulse dipolar electron-spin resonance spectroscopy from the CTT nitroxide to the SQ shows that flavin photoreduction shifts the CTT ~1 nm and increases its motion, without causing full displacement from the protein. dCRY engineered to form the neutral SQ serves as a dark-state proxy to reveal that the CTT remains docked when the flavin ring is reduced but uncharged. Substitutions of flavin-proximal His378 promote CTT undocking in the dark or diminish undocking in the light, consistent with molecular dynamics simulations and TIM degradation activity. The His378 variants inform on recognition motifs for dCRY cellular turnover and strategies for developing optogenetic tools. Chandrasekaran et al. engineered the Drosophila circadian clock protein Cryptochrome (dCRY) to form the neutral semiquinone, which serves as a dark-state proxy. They find that the C-terminal tail of dCRY remains docked when the flavin ring is reduced but uncharged. dCRY His378 variants provide insights into the recognition motifs for dCRY turnover and strategies for optogenetic tools. |
| تدمد: | 2399-3642 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::ad266eedccf9d38cc96583b249acd56a https://doi.org/10.1038/s42003-021-01766-2 |
| حقوق: | OPEN |
| رقم الأكسشن: | edsair.doi.dedup.....ad266eedccf9d38cc96583b249acd56a |
| قاعدة البيانات: | OpenAIRE |
Find this article in full text from Springer Verlag. | تدمد: | 23993642 |
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