Structural determinants underlying the adduct lifetime in the LOV proteins of Pseudomonas putida
| العنوان: | Structural determinants underlying the adduct lifetime in the LOV proteins of Pseudomonas putida |
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| المؤلفون: | Vladimir Arinkin, Joachim Granzin, Karl-Erich Jaeger, Ulrich Krauss, Dieter Willbold, Renu Batra-Safferling |
| المصدر: | 'FEBS Journal ', vol: 288, pages: 4955-4972 (2021) The FEBS journal 288(16), 4955-4972 (2021). doi:10.1111/febs.15785 |
| سنة النشر: | 2021 |
| مصطلحات موضوعية: | 0301 basic medicine, Protein Conformation, Flavin group, Molecular Dynamics Simulation, Biochemistry, Adduct, 03 medical and health sciences, 0302 clinical medicine, Bacterial Proteins, PAS domain, ddc:610, Molecular Biology, chemistry.chemical_classification, biology, Pseudomonas putida, Cell Biology, Protein superfamily, Chromophore, Photochemical Processes, biology.organism_classification, Amino acid, Oxygen, 030104 developmental biology, chemistry, 030220 oncology & carcinogenesis, Helix, Biophysics |
| الوصف: | The primary photochemistry is similar among the flavin-bound sensory domains of light–oxygen–voltage (LOV) photoreceptors, where upon blue-light illumination a covalent adduct is formed on the microseconds time scale between the flavin chromophore and a strictly conserved cysteine residue. In contrast, the adduct-state decay kinetics vary from seconds to days or longer. The molecular basis for this variation among structurally conserved LOV domains is not fully understood. Here, we selected PpSB2-LOV, a fast-cycling (τrec 3.5 min, 20 °C) short LOV protein from Pseudomonas putida that shares 67% sequence identity with a slow-cycling (τrec 2467 min, 20 °C) homologous protein PpSB1-LOV. Based on the crystal structure of the PpSB2-LOV in the dark state reported here, we used a comparative approach, in which we combined structure and sequence information with molecular dynamic (MD) simulations to address the mechanistic basis for the vastly different adduct-state lifetimes in the two homologous proteins. MD simulations pointed toward dynamically distinct structural region, which were subsequently targeted by site-directed mutagenesis of PpSB2-LOV, where we introduced single- and multisite substitutions exchanging them with the corresponding residues from PpSB1-LOV. Collectively, the data presented identify key amino acids on the Aβ-Bβ, Eα-Fα loops, and the Fα helix, such as E27 and I66, that play a decisive role in determining the adduct lifetime. Our results additionally suggest a correlation between the solvent accessibility of the chromophore pocket and adduct-state lifetime. The presented results add to our understanding of LOV signaling and will have important implications in tuning the signaling behavior (on/off kinetics) of LOV-based optogenetic tools. |
| وصف الملف: | application/pdf |
| اللغة: | English |
| تدمد: | 1742-464X |
| الوصول الحر: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::b5a0d0e1ea2406d85596917d4c78631bTest https://epn-library.esrf.fr/flora/jsp/index_view_direct_anonymous.jsp?record=doc:PUB_ESRF:58012Test |
| حقوق: | OPEN |
| رقم الانضمام: | edsair.doi.dedup.....b5a0d0e1ea2406d85596917d4c78631b |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 1742464X |
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