Melanopsin is a photopigment belonging to the G Protein-Coupled Receptor (GPCR) family expressed in a subset of intrinsically photosensitive retinal ganglion cells (ipRGCs) and responsible for a variety of processes. The bistability and, thus, the possibility to function under low retinal availability would make melanopsin a powerful optogenetic tool. Here, we aim to utilize mouse melanopsin to trigger macrophage migration by its subcellular optical activation with localized blue light, while simultaneously imaging the migration with red light. To reduce melanopsin’s red light sensitivity, we employ a combination of in silico structure prediction and automated quantum mechanics/molecular mechanics modeling to predict minimally invasive mutations to shift its absorption spectrum towards the shorter wavelength region of the visible spectrum without compromising the signaling efficiency. The results demonstrate that it is possible to achieve melanopsin mutants that resist red light-induced activation but are activated by blue light and display properties indicating preserved bistability. Using the A333T mutant, we show that the blue light-induced subcellular melanopsin activation triggers localized PIP3 generation and macrophage migration, which we imaged using red light, demonstrating the optogenetic utility of minimally engineered melanopsins.

In-silico predicted mouse melanopsins with blue spectral shifts deliver efficient subcellular signaling / Wijayaratna, D.; Sacchetta, F.; Pedraza-Gonzalez, L.; Fanelli, F.; Sugihara, T.; Koyanagi, M.; Piyawardana, S.; Ghotra, K.; Thotamune, W.; Terakita, A.; Olivucci, M.; Karunarathne, A.. - In: CELL COMMUNICATION AND SIGNALING. - ISSN 1478-811X. - 22:1(2024), pp. 394-394. [10.1186/s12964-024-01753-0]

In-silico predicted mouse melanopsins with blue spectral shifts deliver efficient subcellular signaling

Fanelli F.;
2024

Abstract

Melanopsin is a photopigment belonging to the G Protein-Coupled Receptor (GPCR) family expressed in a subset of intrinsically photosensitive retinal ganglion cells (ipRGCs) and responsible for a variety of processes. The bistability and, thus, the possibility to function under low retinal availability would make melanopsin a powerful optogenetic tool. Here, we aim to utilize mouse melanopsin to trigger macrophage migration by its subcellular optical activation with localized blue light, while simultaneously imaging the migration with red light. To reduce melanopsin’s red light sensitivity, we employ a combination of in silico structure prediction and automated quantum mechanics/molecular mechanics modeling to predict minimally invasive mutations to shift its absorption spectrum towards the shorter wavelength region of the visible spectrum without compromising the signaling efficiency. The results demonstrate that it is possible to achieve melanopsin mutants that resist red light-induced activation but are activated by blue light and display properties indicating preserved bistability. Using the A333T mutant, we show that the blue light-induced subcellular melanopsin activation triggers localized PIP3 generation and macrophage migration, which we imaged using red light, demonstrating the optogenetic utility of minimally engineered melanopsins.
2024
22
1
394
394
In-silico predicted mouse melanopsins with blue spectral shifts deliver efficient subcellular signaling / Wijayaratna, D.; Sacchetta, F.; Pedraza-Gonzalez, L.; Fanelli, F.; Sugihara, T.; Koyanagi, M.; Piyawardana, S.; Ghotra, K.; Thotamune, W.; Terakita, A.; Olivucci, M.; Karunarathne, A.. - In: CELL COMMUNICATION AND SIGNALING. - ISSN 1478-811X. - 22:1(2024), pp. 394-394. [10.1186/s12964-024-01753-0]
Wijayaratna, D.; Sacchetta, F.; Pedraza-Gonzalez, L.; Fanelli, F.; Sugihara, T.; Koyanagi, M.; Piyawardana, S.; Ghotra, K.; Thotamune, W.; Terakita, A...espandi
File in questo prodotto:
File Dimensione Formato  
Melanopsin_CellCommSig2024.pdf

Open access

Tipologia: Versione pubblicata dall'editore
Dimensione 7.71 MB
Formato Adobe PDF
7.71 MB Adobe PDF Visualizza/Apri
Pubblicazioni consigliate

Licenza Creative Commons
I metadati presenti in IRIS UNIMORE sono rilasciati con licenza Creative Commons CC0 1.0 Universal, mentre i file delle pubblicazioni sono rilasciati con licenza Attribuzione 4.0 Internazionale (CC BY 4.0), salvo diversa indicazione.
In caso di violazione di copyright, contattare Supporto Iris

Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11380/1362370
Citazioni
  • ???jsp.display-item.citation.pmc??? 0
  • Scopus 0
  • ???jsp.display-item.citation.isi??? 0
social impact