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• W2989741402 endingPage "1032" @default.
• W2989741402 startingPage "1024" @default.
• W2989741402 abstract "The application of vibrational labels such as thiocyanate (−S−C≡N) for studying protein structure and dynamics is thriving. Absorption spectroscopy is usually employed to obtain wavenumber and line shape of the label. An observable of great significance might be the vibrational lifetime, which can be obtained by pump probe or 2D-IR spectroscopy. Due to the insulating effect of the heavy sulfur atom in the case of the SCN label, the lifetime of the C≡N oscillator is expected to be particularly sensitive to its surrounding as it is not dominated by through-bond relaxation. We therefore investigate the vibrational lifetime of the SCN label at various positions in the blue light sensor protein Photoactive Yellow Protein (PYP) in the ground state and signaling state of the photoreceptor. We find that the vibrational lifetime of the C≡N stretching mode is strongly affected both by its protein environment and by the degree of exposure to the solvent. Even for label positions where the line shape and wavenumber observed by FTIR are barely changing upon activation of the photoreceptor, we find that the lifetime can change considerably. To obtain an unambiguous measure for the solvent exposure of the labeled site, we show that it is imperative to compare the lifetimes in H2O and D2O. Importantly, the lifetimes shorten in H2O as compared to D2O for water exposed labels, while they stay largely the same for buried labels. We quantify this effect by defining a solvent exclusion coefficient (SEC). The response of the label’s vibrational lifetime to its solvent exposure renders it a suitable universal probe for protein investigations. This applies even to systems that are otherwise hard to address, such as transient or short-lived states, which could be created during a protein’s working cycle (as here in PYP) or during protein folding. It is also applicable to flexible systems (intrinsically disordered proteins), protein–protein and protein–membrane interactions." @default.
• W2989741402 created "2019-12-05" @default.
• W2989741402 creator A5004399557 @default.
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• W2989741402 creator A5076681929 @default.
• W2989741402 creator A5077233072 @default.
• W2989741402 date "2019-11-26" @default.
• W2989741402 modified "2023-10-16" @default.
• W2989741402 title "Vibrational Lifetime of the SCN Protein Label in H₂O and D₂O Reports Site-Specific Solvation and Structure Changes During PYP’s Photocycle" @default.
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• W2989741402 doi "https://doi.org/10.1021/acs.analchem.9b03997" @default.
• W2989741402 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/31769286" @default.
• W2989741402 hasPublicationYear "2019" @default.
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