Abstract
During evolution, TRPV1 has lost, retained or selected certain residues at Lipid–Water-Interface (LWI) and formed specific patterns there. The ratio of “hydrophobic–hydrophilic” and “positive–negative-charged” residues at the inner LWI remains conserved throughout vertebrate evolution and plays important role in regulating TRPV1 trafficking and localization. Arg575 is an important residue as Arg575Asp mutant has reduced surface expression, co-localization with lipid raft markers, cell area and increased cell lethality. This lethality is most likely due to the disruption of the ratio between positive–negative charges caused by the mutation. Such lethality can be rescued by either using TRPV1-specfic inhibitor 5′-IRTX or by restoring the positive–negative charge ratio at that position, i.e. by introducing Asp576Arg mutation in Arg575Asp backbone. We propose that Arg575Asp mutation confers TRPV1 in a “constitutive-open-like” condition. These findings have broader implication in understanding the molecular evolution of thermo-sensitive ion channels and the micro-environments involved in processes that goes erratic in different diseases.
Graphical Abstract
The segment of TRPV1 that is present at the inner lipid–water-interface (LWI) has a specific pattern of amino acid combinations. The overall ratio of +ve charge /−ve charge and the ratio of hydrophobicity/hydrophilicity remain constant throughout the vertebrate evolution (ca 450 million years). This specific pattern is not observed in the outer LWI region of TRPV1.
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Data availability
The datasets generated during and/or analysed during the current study are available from the corresponding author on reasonable request.
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Acknowledgements
Intramural funding from NISER, Bhubaneswar is appreciated. CG acknowledges the support and intellectual input from all the present and former lab members. The authors acknowledge Tathagata Mukherjee for his expertise in flow cytometry.
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CG conceived the idea and designed all the experiments. RD1, D., NT, RD2 and SM performed all in silico analyses. RD1, DV, NT, AK, SM and CG analysed all the in silico results. SS performed all the construct preparation. SS and SM did all the cell culture, sample preparation, all the microscopic experiments, image processing, quantification and their statistical analysis. AT performed some functional experiments related to mutant and wild-type channels. CG wrote the paper. The model has been prepared by CG. All authors contributed towards manuscript editing.
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Saha, S., Mohanta, S., Das, R. et al. Ratio of Hydrophobic–Hydrophilic and Positive–Negative Residues at Lipid–Water-Interface Influences Surface Expression and Channel Gating of TRPV1. J Membrane Biol 255, 319–339 (2022). https://doi.org/10.1007/s00232-022-00243-z
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DOI: https://doi.org/10.1007/s00232-022-00243-z