Your browser doesn't support javascript.
loading
Development of a Dual Reporter System to Simultaneously Visualize Ca2+ Signals and AMPK Activity.
Erdogan, Yusuf C; Pilic, Johannes; Gottschalk, Benjamin; Yigit, Esra N; Zaki, Asal G; Öztürk, Gürkan; Eroglu, Emrah; Okutan, Begüm; Sommer, Nicole G; Weinberg, Annelie M; Schindl, Rainer; Graier, Wolfgang F; Malli, Roland.
Afiliación
  • Erdogan YC; Gottfried Schatz Research Center, Molecular Biology and Biochemistry, Medical University of Graz, Neue Stiftingtalstraße 6, Graz 8010, Austria.
  • Pilic J; BioTechMed Graz, Mozartgasse 12/2, Graz 8010, Austria.
  • Gottschalk B; Gottfried Schatz Research Center, Molecular Biology and Biochemistry, Medical University of Graz, Neue Stiftingtalstraße 6, Graz 8010, Austria.
  • Yigit EN; Gottfried Schatz Research Center, Molecular Biology and Biochemistry, Medical University of Graz, Neue Stiftingtalstraße 6, Graz 8010, Austria.
  • Zaki AG; Regenerative and Restorative Medicine Research Center (REMER), Research Institute for Health Sciences and Technologies (SABITA), Istanbul Medipol University, Istanbul 34810, Turkey.
  • Öztürk G; Department of Physiology, International School of Medicine, Istanbul Medipol University, Istanbul 34810, Türkiye.
  • Eroglu E; Regenerative and Restorative Medicine Research Center (REMER), Research Institute for Health Sciences and Technologies (SABITA), Istanbul Medipol University, Istanbul 34810, Turkey.
  • Okutan B; Regenerative and Restorative Medicine Research Center (REMER), Research Institute for Health Sciences and Technologies (SABITA), Istanbul Medipol University, Istanbul 34810, Turkey.
  • Sommer NG; Regenerative and Restorative Medicine Research Center (REMER), Research Institute for Health Sciences and Technologies (SABITA), Istanbul Medipol University, Istanbul 34810, Turkey.
  • Weinberg AM; Department of Orthopedics and Traumatology, Medical University of Graz, Auenbruggerplatz 5, Graz 8036, Austria.
  • Schindl R; Department of Orthopedics and Traumatology, Medical University of Graz, Auenbruggerplatz 5, Graz 8036, Austria.
  • Graier WF; Department of Orthopedics and Traumatology, Medical University of Graz, Auenbruggerplatz 5, Graz 8036, Austria.
  • Malli R; Gottfried Schatz Research Center, Biophysics, Medical University of Graz, Neue Stiftingtalstrasse 6, Graz 8010, Austria.
ACS Sens ; 2024 Aug 21.
Article en En | MEDLINE | ID: mdl-39167044
ABSTRACT
In this study, we introduce a new separation of phases-based activity reporter of kinase (SPARK) for AMP-activated kinase (AMPK), named AMPK-SPARK, which reports the AMPK activation by forming bright fluorescent clusters. Furthermore, we introduce a dual reporter system, named GCaMP-AMPK-SPARK, by incorporating a single-fluorescent protein (FP)-based Ca2+ biosensor, GCaMP6f, into our initial design, enabling simultaneous monitoring of Ca2+ levels and AMPK activity. This system offers the essential quality of information by single-channel fluorescence microscopy without the need for coexpression of different biosensors and elaborate filter layouts to overcome spectral limitations. We used AMPK-SPARK to map endogenous AMPK activity in different cell types and visualized the dynamics of AMPK activation in response to various stimuli. Using GCaMP-AMPK-SPARK, we revealed cell-to-cell heterogeneities in AMPK activation by Ca2+ mobilization. We anticipate that this dual reporter strategy can be employed to study the intricate interplays between different signaling networks and kinase activities.
Palabras clave
Texto completo
Añadir a Mi BVS
Imprimir
XML
PubMed Links
Buscar en Google

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Idioma: En Revista: ACS Sens Año: 2024 Tipo del documento: Article País de afiliación: Austria Pais de publicación: Estados Unidos
Texto completo
Añadir a Mi BVS
Imprimir
XML
PubMed Links
Buscar en Google

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Idioma: En Revista: ACS Sens Año: 2024 Tipo del documento: Article País de afiliación: Austria Pais de publicación: Estados Unidos