Identification and characterisation of Whirlin as a novel modulator of TRPV1
TRPV1 receptor is a molecular transducer for both chemical and physical stimuli in high-threshold afferent neurons (or nociceptors), where it is abundantly expressed and plays an essential role in pain physiopathology. TRPV1 is known to interact with different proteins which modulate receptor activi...
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| Tipo de recurso: | tesis doctoral |
| Fecha de publicación: | 2016 |
| País: | España |
| Institución: | Universidad Miguel Hernández de Elche |
| Repositorio: | REDIUMH. Depósito Digital de la UMH |
| OAI Identifier: | oai:dspace.umh.es:11000/2645 |
| Acceso en línea: | http://hdl.handle.net/11000/2645 |
| Access Level: | acceso abierto |
| Palabra clave: | Bioquímica molecular CDU:5 - Ciencias puras y naturales:57 - Biología:576 - Biología celular y subcelular. Citología CDU:5 - Ciencias puras y naturales:57 - Biología:577 - Bioquímica. Biología molecular. Biofísica |
| Sumario: | TRPV1 receptor is a molecular transducer for both chemical and physical stimuli in high-threshold afferent neurons (or nociceptors), where it is abundantly expressed and plays an essential role in pain physiopathology. TRPV1 is known to interact with different proteins which modulate receptor activity and/or trafficking to the membrane. The formation of these multiproteic complexes, called signalplexes or transducisomes, can alter the activity and/or activation thresholds of TRPV1 receptor and it is crucial for the regulation of channel function. For this reason, the identification of new components of these signalling complexes would allow a more complete understanding of TRPV1 physiopathology, knowledge essential for the development of new pharmacological treatments. In this context, the present study describes the interaction between TRPV1 and Whirlin, a cytosolic protein with PDZ domains previously identified through a yeast two hybrid assay by using the N-terminus of TRPV1 as bait. Specifically, the heterologous expression of Whirlin increases TRPV1 protein levels and promotes the clustering of the receptor to the plasma membrane. Whirlin downregulation through specific silencing RNA results in the concomitant degradation of TRPV1 receptor, which can be prevented by proteasome inhibition. Furthermore, the degradation kinetics of TRPV1 upon arresting protein translation mirrored that of Whirlin in cells co-expressing both proteins, suggesting a parallel degradation mechanism. Finally, Whirlin coexpression attenuated TRPV1 internalisation upon prolonged exposure to the agonist capsaicin. Together, these findings indicate that the assembly of both proteins in a same complex stabilises TRPV1 expression and localisation in the plasma membrane, suggesting that the pharmacological perturbation of this macromolecular complex may represent a novel therapeutic strategy for pain management. |
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