Transient Receptor Potential Vanilloid 1 Signaling Is Independent on Protein Kinase A Phosphorylation of Ankyrin-Rich Membrane Spanning Protein
The sensory ion channel transient receptor potential vanilloid 1 (TRPV1) is mainly expressed in small to medium sized dorsal root ganglion neurons, which are involved in the transfer of acute noxious thermal and chemical stimuli. The Ankyrin-rich membrane spanning protein (ARMS) interaction with TRP...
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MDPI AG
2022-11-01
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| Series: | Medical Sciences |
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| Online Access: | https://www.mdpi.com/2076-3271/10/4/63 |
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| author | Antonio Pellegrino Sandra Mükusch Viola Seitz Christoph Stein Friedrich W. Herberg Harald Seitz |
| author_facet | Antonio Pellegrino Sandra Mükusch Viola Seitz Christoph Stein Friedrich W. Herberg Harald Seitz |
| author_sort | Antonio Pellegrino |
| collection | DOAJ |
| description | The sensory ion channel transient receptor potential vanilloid 1 (TRPV1) is mainly expressed in small to medium sized dorsal root ganglion neurons, which are involved in the transfer of acute noxious thermal and chemical stimuli. The Ankyrin-rich membrane spanning protein (ARMS) interaction with TRPV1 is modulated by protein kinase A (PKA) mediating sensitization. Here, we hypothesize that PKA phosphorylation sites of ARMS are crucial for the modulation of TRPV1 function, and that the phosphorylation of ARMS is facilitated by the A-kinase anchoring protein 79 (AKAP79). We used transfected HEK293 cells, immunoprecipitation, calcium flux, and patch clamp experiments to investigate potential PKA phosphorylation sites in ARMS and in ARMS-related peptides. Additionally, experiments were done to discriminate between PKA and protein kinase D (PKD) phosphorylation. We found different interaction ratios for TRPV1 and ARMS mutants lacking PKA phosphorylation sites. The degree of TRPV1 sensitization by ARMS mutants is independent on PKA phosphorylation. AKAP79 was also involved in the TRPV1/ARMS/PKA signaling complex. These data show that ARMS is a PKA substrate via AKAP79 in the TRPV1 signaling complex and that all four proteins interact physically, regulating TRPV1 sensitization in transfected HEK293 cells. To assess the physiological and/or therapeutic significance of these findings, similar investigations need to be performed in native neurons and/or in vivo. |
| first_indexed | 2024-03-09T16:07:07Z |
| format | Article |
| id | doaj.art-d1350019d9ae443eb9975b4fd90a1a70 |
| institution | Directory Open Access Journal |
| issn | 2076-3271 |
| language | English |
| last_indexed | 2024-03-09T16:07:07Z |
| publishDate | 2022-11-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Medical Sciences |
| spelling | doaj.art-d1350019d9ae443eb9975b4fd90a1a702023-11-24T16:34:33ZengMDPI AGMedical Sciences2076-32712022-11-011046310.3390/medsci10040063Transient Receptor Potential Vanilloid 1 Signaling Is Independent on Protein Kinase A Phosphorylation of Ankyrin-Rich Membrane Spanning ProteinAntonio Pellegrino0Sandra Mükusch1Viola Seitz2Christoph Stein3Friedrich W. Herberg4Harald Seitz5Fraunhofer Institute for Cell Therapy and Immunology, 14476 Potsdam, GermanyFraunhofer Institute for Cell Therapy and Immunology, 14476 Potsdam, GermanyInstitute of Experimental Anaesthesiology, Charité—Universitätsmedizin Berlin, 12203 Berlin, GermanyInstitute of Experimental Anaesthesiology, Charité—Universitätsmedizin Berlin, 12203 Berlin, GermanyDepartment of Biochemistry, University of Kassel, 34132 Kassel, GermanyFraunhofer Institute for Cell Therapy and Immunology, 14476 Potsdam, GermanyThe sensory ion channel transient receptor potential vanilloid 1 (TRPV1) is mainly expressed in small to medium sized dorsal root ganglion neurons, which are involved in the transfer of acute noxious thermal and chemical stimuli. The Ankyrin-rich membrane spanning protein (ARMS) interaction with TRPV1 is modulated by protein kinase A (PKA) mediating sensitization. Here, we hypothesize that PKA phosphorylation sites of ARMS are crucial for the modulation of TRPV1 function, and that the phosphorylation of ARMS is facilitated by the A-kinase anchoring protein 79 (AKAP79). We used transfected HEK293 cells, immunoprecipitation, calcium flux, and patch clamp experiments to investigate potential PKA phosphorylation sites in ARMS and in ARMS-related peptides. Additionally, experiments were done to discriminate between PKA and protein kinase D (PKD) phosphorylation. We found different interaction ratios for TRPV1 and ARMS mutants lacking PKA phosphorylation sites. The degree of TRPV1 sensitization by ARMS mutants is independent on PKA phosphorylation. AKAP79 was also involved in the TRPV1/ARMS/PKA signaling complex. These data show that ARMS is a PKA substrate via AKAP79 in the TRPV1 signaling complex and that all four proteins interact physically, regulating TRPV1 sensitization in transfected HEK293 cells. To assess the physiological and/or therapeutic significance of these findings, similar investigations need to be performed in native neurons and/or in vivo.https://www.mdpi.com/2076-3271/10/4/63TRPV1ARMSPKAAKAP79painphosphorylation |
| spellingShingle | Antonio Pellegrino Sandra Mükusch Viola Seitz Christoph Stein Friedrich W. Herberg Harald Seitz Transient Receptor Potential Vanilloid 1 Signaling Is Independent on Protein Kinase A Phosphorylation of Ankyrin-Rich Membrane Spanning Protein Medical Sciences TRPV1 ARMS PKA AKAP79 pain phosphorylation |
| title | Transient Receptor Potential Vanilloid 1 Signaling Is Independent on Protein Kinase A Phosphorylation of Ankyrin-Rich Membrane Spanning Protein |
| title_full | Transient Receptor Potential Vanilloid 1 Signaling Is Independent on Protein Kinase A Phosphorylation of Ankyrin-Rich Membrane Spanning Protein |
| title_fullStr | Transient Receptor Potential Vanilloid 1 Signaling Is Independent on Protein Kinase A Phosphorylation of Ankyrin-Rich Membrane Spanning Protein |
| title_full_unstemmed | Transient Receptor Potential Vanilloid 1 Signaling Is Independent on Protein Kinase A Phosphorylation of Ankyrin-Rich Membrane Spanning Protein |
| title_short | Transient Receptor Potential Vanilloid 1 Signaling Is Independent on Protein Kinase A Phosphorylation of Ankyrin-Rich Membrane Spanning Protein |
| title_sort | transient receptor potential vanilloid 1 signaling is independent on protein kinase a phosphorylation of ankyrin rich membrane spanning protein |
| topic | TRPV1 ARMS PKA AKAP79 pain phosphorylation |
| url | https://www.mdpi.com/2076-3271/10/4/63 |
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