Proteomic Analysis of the Role of the Adenylyl Cyclase–cAMP Pathway in Red Blood Cell Mechanical Responses
Red blood cell (RBC) deformability is modulated by the phosphorylation status of the cytoskeletal proteins that regulate the interactions of integral transmembrane complexes. Proteomic studies have revealed that receptor-related signaling molecules and regulatory proteins involved in signaling casca...
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MDPI AG
2022-04-01
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| Online Access: | https://www.mdpi.com/2073-4409/11/7/1250 |
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| author | Elif Ugurel Evrim Goksel Neslihan Cilek Elif Kaga Ozlem Yalcin |
| author_facet | Elif Ugurel Evrim Goksel Neslihan Cilek Elif Kaga Ozlem Yalcin |
| author_sort | Elif Ugurel |
| collection | DOAJ |
| description | Red blood cell (RBC) deformability is modulated by the phosphorylation status of the cytoskeletal proteins that regulate the interactions of integral transmembrane complexes. Proteomic studies have revealed that receptor-related signaling molecules and regulatory proteins involved in signaling cascades are present in RBCs. In this study, we investigated the roles of the cAMP signaling mechanism in modulating shear-induced RBC deformability and examined changes in the phosphorylation of the RBC proteome. We implemented the inhibitors of adenylyl cyclase (SQ22536), protein kinase A (H89), and phosphodiesterase (PDE) (pentoxifylline) to whole blood samples, applied 5 Pa shear stress (SS) for 300 s with a capillary tubing system, and evaluated RBC deformability using a LORRCA MaxSis. The inhibition of signaling molecules significantly deteriorated shear-induced RBC deformability (<i>p</i> < 0.05). Capillary SS slightly increased the phosphorylation of RBC cytoskeletal proteins. Tyrosine phosphorylation was significantly elevated by the modulation of the cAMP/PKA pathway (<i>p</i> < 0.05), while serine phosphorylation significantly decreased as a result of the inhibition of PDE (<i>p</i> < 0.05). AC is the core element of this signaling pathway, and PDE works as a negative feedback mechanism that could have potential roles in SS-induced RBC deformability. The cAMP/PKA pathway could regulate RBC deformability during capillary transit by triggering significant alterations in the phosphorylation state of RBCs. |
| first_indexed | 2024-03-09T11:59:53Z |
| format | Article |
| id | doaj.art-b3c29e09d8984aaca4e74bc11208680e |
| institution | Directory Open Access Journal |
| issn | 2073-4409 |
| language | English |
| last_indexed | 2024-03-09T11:59:53Z |
| publishDate | 2022-04-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Cells |
| spelling | doaj.art-b3c29e09d8984aaca4e74bc11208680e2023-11-30T23:05:36ZengMDPI AGCells2073-44092022-04-01117125010.3390/cells11071250Proteomic Analysis of the Role of the Adenylyl Cyclase–cAMP Pathway in Red Blood Cell Mechanical ResponsesElif Ugurel0Evrim Goksel1Neslihan Cilek2Elif Kaga3Ozlem Yalcin4Research Center for Translational Medicine (KUTTAM), Koç University, Istanbul 34450, TurkeyResearch Center for Translational Medicine (KUTTAM), Koç University, Istanbul 34450, TurkeyResearch Center for Translational Medicine (KUTTAM), Koç University, Istanbul 34450, TurkeyHealth Application and Research Center, University of Health Sciences, Afyonkarahisar 03100, TurkeyResearch Center for Translational Medicine (KUTTAM), Koç University, Istanbul 34450, TurkeyRed blood cell (RBC) deformability is modulated by the phosphorylation status of the cytoskeletal proteins that regulate the interactions of integral transmembrane complexes. Proteomic studies have revealed that receptor-related signaling molecules and regulatory proteins involved in signaling cascades are present in RBCs. In this study, we investigated the roles of the cAMP signaling mechanism in modulating shear-induced RBC deformability and examined changes in the phosphorylation of the RBC proteome. We implemented the inhibitors of adenylyl cyclase (SQ22536), protein kinase A (H89), and phosphodiesterase (PDE) (pentoxifylline) to whole blood samples, applied 5 Pa shear stress (SS) for 300 s with a capillary tubing system, and evaluated RBC deformability using a LORRCA MaxSis. The inhibition of signaling molecules significantly deteriorated shear-induced RBC deformability (<i>p</i> < 0.05). Capillary SS slightly increased the phosphorylation of RBC cytoskeletal proteins. Tyrosine phosphorylation was significantly elevated by the modulation of the cAMP/PKA pathway (<i>p</i> < 0.05), while serine phosphorylation significantly decreased as a result of the inhibition of PDE (<i>p</i> < 0.05). AC is the core element of this signaling pathway, and PDE works as a negative feedback mechanism that could have potential roles in SS-induced RBC deformability. The cAMP/PKA pathway could regulate RBC deformability during capillary transit by triggering significant alterations in the phosphorylation state of RBCs.https://www.mdpi.com/2073-4409/11/7/1250red blood cell deformabilitycapillary transitshear stressphosphorylationcytoskeletal proteins |
| spellingShingle | Elif Ugurel Evrim Goksel Neslihan Cilek Elif Kaga Ozlem Yalcin Proteomic Analysis of the Role of the Adenylyl Cyclase–cAMP Pathway in Red Blood Cell Mechanical Responses Cells red blood cell deformability capillary transit shear stress phosphorylation cytoskeletal proteins |
| title | Proteomic Analysis of the Role of the Adenylyl Cyclase–cAMP Pathway in Red Blood Cell Mechanical Responses |
| title_full | Proteomic Analysis of the Role of the Adenylyl Cyclase–cAMP Pathway in Red Blood Cell Mechanical Responses |
| title_fullStr | Proteomic Analysis of the Role of the Adenylyl Cyclase–cAMP Pathway in Red Blood Cell Mechanical Responses |
| title_full_unstemmed | Proteomic Analysis of the Role of the Adenylyl Cyclase–cAMP Pathway in Red Blood Cell Mechanical Responses |
| title_short | Proteomic Analysis of the Role of the Adenylyl Cyclase–cAMP Pathway in Red Blood Cell Mechanical Responses |
| title_sort | proteomic analysis of the role of the adenylyl cyclase camp pathway in red blood cell mechanical responses |
| topic | red blood cell deformability capillary transit shear stress phosphorylation cytoskeletal proteins |
| url | https://www.mdpi.com/2073-4409/11/7/1250 |
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