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Interactive computa...
Interactive computational and experimental approaches improve the sensitivity of periplasmic binding protein-based nicotine biosensors for measurements in biofluids
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- Haloi, Nandan (författare)
- KTH,Biofysik,Science for Life Laboratory, SciLifeLab
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- Huang, Shan (författare)
- Division of Biology and Biological Engineering, California Institute of Technology, Pasadena, CA 91125, USA
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- Nichols, Aaron L. (författare)
- Division of Biology and Biological Engineering, California Institute of Technology, Pasadena, CA 91125, USA
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- Fine, Eve J. (författare)
- Division of Biology and Biological Engineering, California Institute of Technology, Pasadena, CA 91125, USA
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- Friesenhahn, Nicholas J. (författare)
- Division of Chemistry and Chemical Engineering, California Institute of Technology, Pasadena, CA 91125, USA
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- Marotta, Christopher B. (författare)
- Division of Chemistry and Chemical Engineering, California Institute of Technology, Pasadena, CA 91125, USA
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- Dougherty, Dennis A. (författare)
- Division of Chemistry and Chemical Engineering, California Institute of Technology, Pasadena, CA 91125, USA
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- Lindahl, Erik, 1972- (författare)
- Stockholms universitet,KTH,Biofysik,Science for Life Laboratory, SciLifeLab,Institutionen för biokemi och biofysik,Science for Life Laboratory (SciLifeLab),KTH Royal Institute of Technology, Sweden
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- Howard, Rebecca J., 1980- (författare)
- Stockholms universitet,Institutionen för biokemi och biofysik,Science for Life Laboratory (SciLifeLab)
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- Mayo, Stephen L. (författare)
- Division of Biology and Biological Engineering, California Institute of Technology, Pasadena, CA 91125, USA
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- Lester, Henry A. (författare)
- Division of Biology and Biological Engineering, California Institute of Technology, Pasadena, CA 91125, USA
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(creator_code:org_t)
- Oxford University Press (OUP), 2024
- 2024
- Engelska.
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Ingår i: Protein Engineering Design & Selection. - : Oxford University Press (OUP). - 1741-0126 .- 1741-0134. ; 37
- Relaterad länk:
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https://urn.kb.se/re...
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https://doi.org/10.1...
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https://urn.kb.se/re...
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Abstract
Ämnesord
Stäng
- We developed fluorescent protein sensors for nicotine with improved sensitivity. For iNicSnFR12 at pH 7.4, the proportionality constant for ∆F/F0 vs [nicotine] (δ-slope, 2.7 μM−1) is 6.1-fold higher than the previously reported iNicSnFR3a. The activated state of iNicSnFR12 has a fluorescence quantum yield of at least 0.6. We measured similar dose-response relations for the nicotine-induced absorbance increase and fluorescence increase, suggesting that the absorbance increase leads to the fluorescence increase via the previously described nicotine-induced conformational change, the ‘candle snuffer’ mechanism. Molecular dynamics (MD) simulations identified a binding pose for nicotine, previously indeterminate from experimental data. MD simulations also showed that Helix 4 of the periplasmic binding protein (PBP) domain appears tilted in iNicSnFR12 relative to iNicSnFR3a, likely altering allosteric network(s) that link the ligand binding site to the fluorophore. In thermal melt experiments, nicotine stabilized the PBP of the tested iNicSnFR variants. iNicSnFR12 resolved nicotine in diluted mouse and human serum at 100 nM, the peak [nicotine] that occurs during smoking or vaping, and possibly at the decreasing levels during intervals between sessions. NicSnFR12 was also partially activated by unidentified endogenous ligand(s) in biofluids. Improved iNicSnFR12 variants could become the molecular sensors in continuous nicotine monitors for animal and human biofluids.
Ämnesord
- NATURVETENSKAP -- Biologi (hsv//swe)
- NATURAL SCIENCES -- Biological Sciences (hsv//eng)
- NATURVETENSKAP -- Biologi -- Biokemi och molekylärbiologi (hsv//swe)
- NATURAL SCIENCES -- Biological Sciences -- Biochemistry and Molecular Biology (hsv//eng)
- NATURVETENSKAP -- Biologi -- Biofysik (hsv//swe)
- NATURAL SCIENCES -- Biological Sciences -- Biophysics (hsv//eng)
Nyckelord
- absorption
- biosensor
- computation
- fluorescence
- iNicSnFR
Publikations- och innehållstyp
- ref (ämneskategori)
- art (ämneskategori)
Hitta via bibliotek
Till lärosätets databas
- Av författaren/redakt...
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Haloi, Nandan
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Huang, Shan
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Nichols, Aaron L ...
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Fine, Eve J.
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Friesenhahn, Nic ...
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Marotta, Christo ...
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visa fler...
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Dougherty, Denni ...
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Lindahl, Erik, 1 ...
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Howard, Rebecca ...
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Mayo, Stephen L.
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Lester, Henry A.
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- Om ämnet
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- NATURVETENSKAP
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NATURVETENSKAP
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och Biologi
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- NATURVETENSKAP
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NATURVETENSKAP
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och Biologi
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och Biokemi och mole ...
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- NATURVETENSKAP
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NATURVETENSKAP
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och Biologi
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och Biofysik
- Artiklar i publikationen
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Protein Engineer ...
- Av lärosätet
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Kungliga Tekniska Högskolan
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Stockholms universitet