Glucose-dependent phosphorylation signaling pathways and crosstalk to mitochondrial respiration in insulin secreting cells
Abstract Background Glucose is the main secretagogue of pancreatic beta-cells. Uptake and metabolism of the nutrient stimulates the beta-cell to release the blood glucose lowering hormone insulin. This metabolic activation is associated with a pronounced increase in mitochondrial respiration. Glucos...
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doaj-630414b375bd4c67a6b87d4f1b051dc92020-11-25T02:52:04ZengBMCCell Communication and Signaling1478-811X2019-02-0117111910.1186/s12964-019-0326-6Glucose-dependent phosphorylation signaling pathways and crosstalk to mitochondrial respiration in insulin secreting cellsJaime Santo-Domingo0Antonio Núñez Galindo1Ornella Cominetti2Umberto De Marchi3Pedro Cutillas4Loïc Dayon5Andreas Wiederkehr6Nestlé Institute of Health Sciences, Nestlé ResearchNestlé Institute of Health Sciences, Nestlé ResearchNestlé Institute of Health Sciences, Nestlé ResearchNestlé Institute of Health Sciences, Nestlé ResearchAnalytical Signalling Group, Centre for Cell Signalling, Queen Mary University of LondonNestlé Institute of Health Sciences, Nestlé ResearchNestlé Institute of Health Sciences, Nestlé ResearchAbstract Background Glucose is the main secretagogue of pancreatic beta-cells. Uptake and metabolism of the nutrient stimulates the beta-cell to release the blood glucose lowering hormone insulin. This metabolic activation is associated with a pronounced increase in mitochondrial respiration. Glucose stimulation also initiates a number of signal transduction pathways for the coordinated regulation of multiple biological processes required for insulin secretion. Methods Shotgun proteomics including TiO2 enrichment of phosphorylated peptides followed by liquid chromatography tandem mass spectrometry on lysates from glucose-stimulated INS-1E cells was used to identify glucose regulated phosphorylated proteins and signal transduction pathways. Kinase substrate enrichment analysis (KSEA) was applied to identify key regulated kinases and phosphatases. Glucose-induced oxygen consumption was measured using a XF96 Seahorse instrument to reveal cross talk between glucose-regulated kinases and mitochondrial activation. Results Our kinetic analysis of substrate phosphorylation reveal the molecular mechanism leading to rapid activation of insulin biogenesis, vesicle trafficking, insulin granule exocytosis and cytoskeleton remodeling. Kinase-substrate enrichment identified upstream kinases and phosphatases and time-dependent activity changes during glucose stimulation. Activity trajectories of well-known glucose-regulated kinases and phosphatases are described. In addition, we predict activity changes in a number of kinases including NUAK1, not or only poorly studied in the context of the pancreatic beta-cell. Furthermore, we pharmacologically tested whether signaling pathways predicted by kinase-substrate enrichment analysis affected glucose-dependent acceleration of mitochondrial respiration. We find that phosphoinositide 3-kinase, Ca2+/calmodulin dependent protein kinase and protein kinase C contribute to short-term regulation of energy metabolism. Conclusions Our results provide a global view into the regulation of kinases and phosphatases in insulin secreting cells and suggest cross talk between glucose-induced signal transduction and mitochondrial activation.http://link.springer.com/article/10.1186/s12964-019-0326-6GlucoseMass spectrometryMetabolismMitochondriaSignalingPhospho-proteome |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Jaime Santo-Domingo Antonio Núñez Galindo Ornella Cominetti Umberto De Marchi Pedro Cutillas Loïc Dayon Andreas Wiederkehr |
spellingShingle |
Jaime Santo-Domingo Antonio Núñez Galindo Ornella Cominetti Umberto De Marchi Pedro Cutillas Loïc Dayon Andreas Wiederkehr Glucose-dependent phosphorylation signaling pathways and crosstalk to mitochondrial respiration in insulin secreting cells Cell Communication and Signaling Glucose Mass spectrometry Metabolism Mitochondria Signaling Phospho-proteome |
author_facet |
Jaime Santo-Domingo Antonio Núñez Galindo Ornella Cominetti Umberto De Marchi Pedro Cutillas Loïc Dayon Andreas Wiederkehr |
author_sort |
Jaime Santo-Domingo |
title |
Glucose-dependent phosphorylation signaling pathways and crosstalk to mitochondrial respiration in insulin secreting cells |
title_short |
Glucose-dependent phosphorylation signaling pathways and crosstalk to mitochondrial respiration in insulin secreting cells |
title_full |
Glucose-dependent phosphorylation signaling pathways and crosstalk to mitochondrial respiration in insulin secreting cells |
title_fullStr |
Glucose-dependent phosphorylation signaling pathways and crosstalk to mitochondrial respiration in insulin secreting cells |
title_full_unstemmed |
Glucose-dependent phosphorylation signaling pathways and crosstalk to mitochondrial respiration in insulin secreting cells |
title_sort |
glucose-dependent phosphorylation signaling pathways and crosstalk to mitochondrial respiration in insulin secreting cells |
publisher |
BMC |
series |
Cell Communication and Signaling |
issn |
1478-811X |
publishDate |
2019-02-01 |
description |
Abstract Background Glucose is the main secretagogue of pancreatic beta-cells. Uptake and metabolism of the nutrient stimulates the beta-cell to release the blood glucose lowering hormone insulin. This metabolic activation is associated with a pronounced increase in mitochondrial respiration. Glucose stimulation also initiates a number of signal transduction pathways for the coordinated regulation of multiple biological processes required for insulin secretion. Methods Shotgun proteomics including TiO2 enrichment of phosphorylated peptides followed by liquid chromatography tandem mass spectrometry on lysates from glucose-stimulated INS-1E cells was used to identify glucose regulated phosphorylated proteins and signal transduction pathways. Kinase substrate enrichment analysis (KSEA) was applied to identify key regulated kinases and phosphatases. Glucose-induced oxygen consumption was measured using a XF96 Seahorse instrument to reveal cross talk between glucose-regulated kinases and mitochondrial activation. Results Our kinetic analysis of substrate phosphorylation reveal the molecular mechanism leading to rapid activation of insulin biogenesis, vesicle trafficking, insulin granule exocytosis and cytoskeleton remodeling. Kinase-substrate enrichment identified upstream kinases and phosphatases and time-dependent activity changes during glucose stimulation. Activity trajectories of well-known glucose-regulated kinases and phosphatases are described. In addition, we predict activity changes in a number of kinases including NUAK1, not or only poorly studied in the context of the pancreatic beta-cell. Furthermore, we pharmacologically tested whether signaling pathways predicted by kinase-substrate enrichment analysis affected glucose-dependent acceleration of mitochondrial respiration. We find that phosphoinositide 3-kinase, Ca2+/calmodulin dependent protein kinase and protein kinase C contribute to short-term regulation of energy metabolism. Conclusions Our results provide a global view into the regulation of kinases and phosphatases in insulin secreting cells and suggest cross talk between glucose-induced signal transduction and mitochondrial activation. |
topic |
Glucose Mass spectrometry Metabolism Mitochondria Signaling Phospho-proteome |
url |
http://link.springer.com/article/10.1186/s12964-019-0326-6 |
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