FLIM‐FRET‐based analysis of S100A11/annexin interactions in living cells
Proteins achieve their biological functions in cells by cooperation in protein complexes. In this study, we employed fluorescence lifetime imaging microscopy (FLIM)‐based Förster resonance energy transfer (FRET) measurements to investigate protein complexes comprising S100A11 and different members o...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Wiley
2024-04-01
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| Series: | FEBS Open Bio |
| Subjects: | |
| Online Access: | https://doi.org/10.1002/2211-5463.13782 |
| _version_ | 1827294684893937664 |
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| author | Christian Melle Birgit Hoffmann Annett Wiesenburg Christoph Biskup |
| author_facet | Christian Melle Birgit Hoffmann Annett Wiesenburg Christoph Biskup |
| author_sort | Christian Melle |
| collection | DOAJ |
| description | Proteins achieve their biological functions in cells by cooperation in protein complexes. In this study, we employed fluorescence lifetime imaging microscopy (FLIM)‐based Förster resonance energy transfer (FRET) measurements to investigate protein complexes comprising S100A11 and different members of the annexin (ANX) family, such as ANXA1, ANXA2, ANXA4, ANXA5, and AnxA6, in living cells. Using an S100A11 mutant without the capacity for Ca2+ binding, we found that Ca2+ binding of S100A11 is important for distinct S100A11/ANXA2 complex formation; however, ANXA1‐containing complexes were unaffected by this mutant. An increase in the intracellular calcium concentration induced calcium ionophores, which strengthened the ANXA2/S100A11 interaction. Furthermore, we were able to show that S100A11 also interacts with ANXA4 in living cells. The FLIM‐FRET approach used here can serve as a tool to analyze interactions between S100A11 and distinct annexins under physiological conditions in living cells. |
| first_indexed | 2024-04-24T14:06:19Z |
| format | Article |
| id | doaj.art-2dbc77c3822943059cfd9ac8fc8a30d4 |
| institution | Directory Open Access Journal |
| issn | 2211-5463 |
| language | English |
| last_indexed | 2024-04-24T14:06:19Z |
| publishDate | 2024-04-01 |
| publisher | Wiley |
| record_format | Article |
| series | FEBS Open Bio |
| spelling | doaj.art-2dbc77c3822943059cfd9ac8fc8a30d42024-04-03T10:12:36ZengWileyFEBS Open Bio2211-54632024-04-0114462664210.1002/2211-5463.13782FLIM‐FRET‐based analysis of S100A11/annexin interactions in living cellsChristian Melle0Birgit Hoffmann1Annett Wiesenburg2Christoph Biskup3Biomolecular Photonics Group, Jena University Hospital Friedrich Schiller University Jena GermanyBiomolecular Photonics Group, Jena University Hospital Friedrich Schiller University Jena GermanyBiomolecular Photonics Group, Jena University Hospital Friedrich Schiller University Jena GermanyBiomolecular Photonics Group, Jena University Hospital Friedrich Schiller University Jena GermanyProteins achieve their biological functions in cells by cooperation in protein complexes. In this study, we employed fluorescence lifetime imaging microscopy (FLIM)‐based Förster resonance energy transfer (FRET) measurements to investigate protein complexes comprising S100A11 and different members of the annexin (ANX) family, such as ANXA1, ANXA2, ANXA4, ANXA5, and AnxA6, in living cells. Using an S100A11 mutant without the capacity for Ca2+ binding, we found that Ca2+ binding of S100A11 is important for distinct S100A11/ANXA2 complex formation; however, ANXA1‐containing complexes were unaffected by this mutant. An increase in the intracellular calcium concentration induced calcium ionophores, which strengthened the ANXA2/S100A11 interaction. Furthermore, we were able to show that S100A11 also interacts with ANXA4 in living cells. The FLIM‐FRET approach used here can serve as a tool to analyze interactions between S100A11 and distinct annexins under physiological conditions in living cells.https://doi.org/10.1002/2211-5463.13782annexinFLIMFRETprotein–protein interactionsS100 proteinS100A11 |
| spellingShingle | Christian Melle Birgit Hoffmann Annett Wiesenburg Christoph Biskup FLIM‐FRET‐based analysis of S100A11/annexin interactions in living cells FEBS Open Bio annexin FLIM FRET protein–protein interactions S100 protein S100A11 |
| title | FLIM‐FRET‐based analysis of S100A11/annexin interactions in living cells |
| title_full | FLIM‐FRET‐based analysis of S100A11/annexin interactions in living cells |
| title_fullStr | FLIM‐FRET‐based analysis of S100A11/annexin interactions in living cells |
| title_full_unstemmed | FLIM‐FRET‐based analysis of S100A11/annexin interactions in living cells |
| title_short | FLIM‐FRET‐based analysis of S100A11/annexin interactions in living cells |
| title_sort | flim fret based analysis of s100a11 annexin interactions in living cells |
| topic | annexin FLIM FRET protein–protein interactions S100 protein S100A11 |
| url | https://doi.org/10.1002/2211-5463.13782 |
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