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Interactive computational and experimental approaches improve the sensitivity of periplasmic binding protein-based nicotine biosensors for measurements in biofluids.
bioRxiv. 2023 Aug 07B

Abstract

We developed fluorescent protein sensors for nicotine with improved sensitivity. For iNicSnFR12 at pH 7.4, ΔF/F0 increased with nicotine concentration; the proportionality constant (S-slope), 2.6 μM[-1], is 6.5-fold higher than the previously reported iNicSnFR3a. Fluorescence activation results primarily via increased absorption. We identified a binding pose for nicotine, previously indeterminate from experimental data. Helix 4 appears tilted in iNicSnFR12 relative to iNicSnFR3a, likely altering allosteric network(s) that link the ligand binding site to the fluorophore. Nicotine stabilizes the PBP domains of the tested iNicSnFR variants. iNicSnFR12 resolved nicotine in diluted mouse and human serum at 100 nM, the peak concentration that occurs during smoking or vaping, and possibly at the decreasing levels during intervals between sessions. NicSnFR12 is also partially activated by unidentified endogenous ligand(s) in biofluids. iNicSnFR12 or an improved variant could become the molecular sensor in a continuous nicotine monitor for animal and human biofluids.

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Authors+Show Affiliations

Haloi N
Department of Applied Physics, Science for Life Laboratory, KTH Royal Institute of Technology, 10044 Stockholm, Sweden.
Huang S
Division of Biology and Biological Engineering, California Institute of Technology, Pasadena CA 91125 USA.
Nichols AL
Division of Biology and Biological Engineering, California Institute of Technology, Pasadena CA 91125 USA.
Fine EJ
Division of Biology and Biological Engineering, California Institute of Technology, Pasadena CA 91125 USA.
Friesenhahn NJ
Division of Chemistry and Chemical Engineering, California Institute of Technology, Pasadena CA 91125 USA.
Marotta CB
Division of Chemistry and Chemical Engineering, California Institute of Technology, Pasadena CA 91125 USA.
Dougherty DA
Division of Chemistry and Chemical Engineering, California Institute of Technology, Pasadena CA 91125 USA.
Lindahl E
Department of Applied Physics, Science for Life Laboratory, KTH Royal Institute of Technology, 10044 Stockholm, Sweden. Department of Biochemistry and Biophysics, Science for Life Laboratory, Stockholm University, 10691 Stockholm, Sweden.
Howard RJ
Department of Biochemistry and Biophysics, Science for Life Laboratory, Stockholm University, 10691 Stockholm, Sweden.
Mayo SL
Division of Biology and Biological Engineering, California Institute of Technology, Pasadena CA 91125 USA.
Lester HA
Division of Biology and Biological Engineering, California Institute of Technology, Pasadena CA 91125 USA.

Pub Type(s)

Preprint

Language

eng

PubMed ID

36712031

Citation

Haloi, Nandan, et al. "Interactive Computational and Experimental Approaches Improve the Sensitivity of Periplasmic Binding Protein-based Nicotine Biosensors for Measurements in Biofluids." BioRxiv : the Preprint Server for Biology, 2023.
Haloi N, Huang S, Nichols AL, et al. Interactive computational and experimental approaches improve the sensitivity of periplasmic binding protein-based nicotine biosensors for measurements in biofluids. bioRxiv. 2023.
Haloi, N., Huang, S., Nichols, A. L., Fine, E. J., Friesenhahn, N. J., Marotta, C. B., Dougherty, D. A., Lindahl, E., Howard, R. J., Mayo, S. L., & Lester, H. A. (2023). Interactive computational and experimental approaches improve the sensitivity of periplasmic binding protein-based nicotine biosensors for measurements in biofluids. BioRxiv : the Preprint Server for Biology. https://doi.org/10.1101/2023.01.16.524298
Haloi N, et al. Interactive Computational and Experimental Approaches Improve the Sensitivity of Periplasmic Binding Protein-based Nicotine Biosensors for Measurements in Biofluids. bioRxiv. 2023 Aug 7; PubMed PMID: 36712031.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Interactive computational and experimental approaches improve the sensitivity of periplasmic binding protein-based nicotine biosensors for measurements in biofluids. AU - Haloi,Nandan, AU - Huang,Shan, AU - Nichols,Aaron L, AU - Fine,Eve J, AU - Friesenhahn,Nicholas J, AU - Marotta,Christopher B, AU - Dougherty,Dennis A, AU - Lindahl,Erik, AU - Howard,Rebecca J, AU - Mayo,Stephen L, AU - Lester,Henry A, Y1 - 2023/08/07/ PY - 2023/1/31/pubmed PY - 2023/1/31/medline PY - 2023/1/30/entrez JF - bioRxiv : the preprint server for biology JO - bioRxiv N2 - We developed fluorescent protein sensors for nicotine with improved sensitivity. For iNicSnFR12 at pH 7.4, ΔF/F0 increased with nicotine concentration; the proportionality constant (S-slope), 2.6 μM[-1], is 6.5-fold higher than the previously reported iNicSnFR3a. Fluorescence activation results primarily via increased absorption. We identified a binding pose for nicotine, previously indeterminate from experimental data. Helix 4 appears tilted in iNicSnFR12 relative to iNicSnFR3a, likely altering allosteric network(s) that link the ligand binding site to the fluorophore. Nicotine stabilizes the PBP domains of the tested iNicSnFR variants. iNicSnFR12 resolved nicotine in diluted mouse and human serum at 100 nM, the peak concentration that occurs during smoking or vaping, and possibly at the decreasing levels during intervals between sessions. NicSnFR12 is also partially activated by unidentified endogenous ligand(s) in biofluids. iNicSnFR12 or an improved variant could become the molecular sensor in a continuous nicotine monitor for animal and human biofluids. UR - https://www.unboundmedicine.com/medline/citation/36712031/Interactive_computational_and_experimental_approaches_improve_the_sensitivity_of_periplasmic_binding_protein_based_nicotine_biosensors_for_measurements_in_biofluids_ DB - PRIME DP - Unbound Medicine ER -