Recombinant Production, NMR Solution Structure, and Membrane Interaction of the Phα1β Toxin, a TRPA1 Modulator from the Brazilian Armed Spider Phoneutria nigriventer
Abstract
:1. Introduction
2. Results and Discussion
2.1. Expression and Purification
2.2. Spatial Structure of Phα1β
2.3. Backbone Dynamics of Phα1β
2.4. Properties of the Phα1β Surface
2.5. Interaction of Phα1β with Lipid Bilayers
2.6. Interaction of Phα1β with Rat TRPA1 Expressed in Xenopus Oocytes
3. Conclusions
4. Materials and Methods
4.1. Bacterial Expression of Phα1β
4.2. Liquid Chromatography and Mass Spectrometry
4.3. NMR Experiments and Spatial Structure Calculation
4.4. Liposome Titration
4.5. Electrophysiological Recordings
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Lipids | Partition Equilibrium a Kp | Langmuir Isotherm b | |
---|---|---|---|
KN | N | ||
×103·M−1 | ×106·M−1 | ||
POPC/POPG (3:1) | 5.1 ± 0.1 | 2.8 ± 0.7 | 41 ± 1 |
POPC/POPG (3:1), 150 mM NaCl | 0.20 ± 0.02 | 0.0087 ± 0.0046 | 41 c |
POPC, 150 mM NaCl | 0.14 ± 0.01 | 0.0060 ± 0.0013 | 41 c |
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Lyukmanova, E.N.; Mironov, P.A.; Kulbatskii, D.S.; Shulepko, M.A.; Paramonov, A.S.; Chernaya, E.M.; Logashina, Y.A.; Andreev, Y.A.; Kirpichnikov, M.P.; Shenkarev, Z.O. Recombinant Production, NMR Solution Structure, and Membrane Interaction of the Phα1β Toxin, a TRPA1 Modulator from the Brazilian Armed Spider Phoneutria nigriventer. Toxins 2023, 15, 378. https://doi.org/10.3390/toxins15060378
Lyukmanova EN, Mironov PA, Kulbatskii DS, Shulepko MA, Paramonov AS, Chernaya EM, Logashina YA, Andreev YA, Kirpichnikov MP, Shenkarev ZO. Recombinant Production, NMR Solution Structure, and Membrane Interaction of the Phα1β Toxin, a TRPA1 Modulator from the Brazilian Armed Spider Phoneutria nigriventer. Toxins. 2023; 15(6):378. https://doi.org/10.3390/toxins15060378
Chicago/Turabian StyleLyukmanova, Ekaterina N., Pavel A. Mironov, Dmitrii S. Kulbatskii, Mikhail A. Shulepko, Alexander S. Paramonov, Elizaveta M. Chernaya, Yulia A. Logashina, Yaroslav A. Andreev, Mikhail P. Kirpichnikov, and Zakhar O. Shenkarev. 2023. "Recombinant Production, NMR Solution Structure, and Membrane Interaction of the Phα1β Toxin, a TRPA1 Modulator from the Brazilian Armed Spider Phoneutria nigriventer" Toxins 15, no. 6: 378. https://doi.org/10.3390/toxins15060378