Searching for protein partners of short-chain 3-hydroxyacyl-CoA dehydrogenase (SCHAD) reveals keratin 8 as a novel candidate for interaction in pancreatic β-cells
Abstract Background Short-chain 3-hydroxyacyl-CoA dehydrogenase (SCHAD) is a ubiquitously expressed mitochondrial enzyme with a role in the degradation of fatty acids. Because the protein also is a negative regulator of insulin secretion in pancreatic β-cells, inactivating mutations in the SCHAD gen...
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BMC
2025-06-01
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| Series: | BMC Molecular and Cell Biology |
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| Online Access: | https://doi.org/10.1186/s12860-025-00544-w |
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| author | Kelly Velasco Janniche Torsvik Johanna L. St-Louis Sarah Baghestani Jonas S. G. Silvander Rohit N. Kulkarni Diana M. Toivola Anders Molven |
| author_facet | Kelly Velasco Janniche Torsvik Johanna L. St-Louis Sarah Baghestani Jonas S. G. Silvander Rohit N. Kulkarni Diana M. Toivola Anders Molven |
| author_sort | Kelly Velasco |
| collection | DOAJ |
| description | Abstract Background Short-chain 3-hydroxyacyl-CoA dehydrogenase (SCHAD) is a ubiquitously expressed mitochondrial enzyme with a role in the degradation of fatty acids. Because the protein also is a negative regulator of insulin secretion in pancreatic β-cells, inactivating mutations in the SCHAD gene (HADH) cause congenital hyperinsulinism of infancy (CHI) and severe hypoglycemia. Here we sought to identify novel interaction partners of SCHAD that might be particularly relevant for the endocrine pancreas. Results Employing the SCHAD protein as bait, we performed yeast 2-hybrid screening of a cDNA library made from human islets of Langerhans. Surprisingly, the screening revealed the intermediate filament protein keratin 8 (K8) as a putative interaction partner of SCHAD with very high confidence. Previous reports have linked K8 to glucose homeostasis, and we confirmed the SCHAD interaction by co-immunoprecipitation in HEK293 cells. SCHAD and K8 expression were then characterized in the human β-cell model EndoC-βH1. By using proximity ligation assay, we demonstrated that stimulating the cells with a high level of glucose triggered a transient increase in the interaction. However, when studying knockout mice, we found that the loss of either K8 or SCHAD did not change the expression level of the other interaction partner. Still, when K8 knockout mice were challenged with a ketogenic diet, upregulation of SCHAD expression was blunted compared to the upregulation observed in wildtype littermates. Conclusions We propose that the SCHAD protein interacts with K8 in a way that might be relevant for proper functioning of the pancreatic β-cell. Whether the SCHAD-K8 interaction influences the phenotype of CHI remains to be demonstrated. |
| format | Article |
| id | doaj-art-191e7317cb234f21b91dc2d3281db458 |
| institution | OA Journals |
| issn | 2661-8850 |
| language | English |
| publishDate | 2025-06-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Molecular and Cell Biology |
| spelling | doaj-art-191e7317cb234f21b91dc2d3281db4582025-08-20T02:05:42ZengBMCBMC Molecular and Cell Biology2661-88502025-06-0126111510.1186/s12860-025-00544-wSearching for protein partners of short-chain 3-hydroxyacyl-CoA dehydrogenase (SCHAD) reveals keratin 8 as a novel candidate for interaction in pancreatic β-cellsKelly Velasco0Janniche Torsvik1Johanna L. St-Louis2Sarah Baghestani3Jonas S. G. Silvander4Rohit N. Kulkarni5Diana M. Toivola6Anders Molven7Gade Laboratory for Pathology, Department of Clinical Medicine, University of BergenGade Laboratory for Pathology, Department of Clinical Medicine, University of BergenGade Laboratory for Pathology, Department of Clinical Medicine, University of BergenCell Biology, Faculty of Science and Engineering, Åbo Akademi UniversityCell Biology, Faculty of Science and Engineering, Åbo Akademi UniversityIslet Cell and Regenerative Biology, Joslin Diabetes Center, Harvard Medical SchoolCell Biology, Faculty of Science and Engineering, Åbo Akademi UniversityGade Laboratory for Pathology, Department of Clinical Medicine, University of BergenAbstract Background Short-chain 3-hydroxyacyl-CoA dehydrogenase (SCHAD) is a ubiquitously expressed mitochondrial enzyme with a role in the degradation of fatty acids. Because the protein also is a negative regulator of insulin secretion in pancreatic β-cells, inactivating mutations in the SCHAD gene (HADH) cause congenital hyperinsulinism of infancy (CHI) and severe hypoglycemia. Here we sought to identify novel interaction partners of SCHAD that might be particularly relevant for the endocrine pancreas. Results Employing the SCHAD protein as bait, we performed yeast 2-hybrid screening of a cDNA library made from human islets of Langerhans. Surprisingly, the screening revealed the intermediate filament protein keratin 8 (K8) as a putative interaction partner of SCHAD with very high confidence. Previous reports have linked K8 to glucose homeostasis, and we confirmed the SCHAD interaction by co-immunoprecipitation in HEK293 cells. SCHAD and K8 expression were then characterized in the human β-cell model EndoC-βH1. By using proximity ligation assay, we demonstrated that stimulating the cells with a high level of glucose triggered a transient increase in the interaction. However, when studying knockout mice, we found that the loss of either K8 or SCHAD did not change the expression level of the other interaction partner. Still, when K8 knockout mice were challenged with a ketogenic diet, upregulation of SCHAD expression was blunted compared to the upregulation observed in wildtype littermates. Conclusions We propose that the SCHAD protein interacts with K8 in a way that might be relevant for proper functioning of the pancreatic β-cell. Whether the SCHAD-K8 interaction influences the phenotype of CHI remains to be demonstrated.https://doi.org/10.1186/s12860-025-00544-wCongenital hyperinsulinism of infancyShort-chain 3-hydroxyacyl-CoA dehydrogenaseSCHADHADHIntermediate filamentsKeratin 8 |
| spellingShingle | Kelly Velasco Janniche Torsvik Johanna L. St-Louis Sarah Baghestani Jonas S. G. Silvander Rohit N. Kulkarni Diana M. Toivola Anders Molven Searching for protein partners of short-chain 3-hydroxyacyl-CoA dehydrogenase (SCHAD) reveals keratin 8 as a novel candidate for interaction in pancreatic β-cells BMC Molecular and Cell Biology Congenital hyperinsulinism of infancy Short-chain 3-hydroxyacyl-CoA dehydrogenase SCHAD HADH Intermediate filaments Keratin 8 |
| title | Searching for protein partners of short-chain 3-hydroxyacyl-CoA dehydrogenase (SCHAD) reveals keratin 8 as a novel candidate for interaction in pancreatic β-cells |
| title_full | Searching for protein partners of short-chain 3-hydroxyacyl-CoA dehydrogenase (SCHAD) reveals keratin 8 as a novel candidate for interaction in pancreatic β-cells |
| title_fullStr | Searching for protein partners of short-chain 3-hydroxyacyl-CoA dehydrogenase (SCHAD) reveals keratin 8 as a novel candidate for interaction in pancreatic β-cells |
| title_full_unstemmed | Searching for protein partners of short-chain 3-hydroxyacyl-CoA dehydrogenase (SCHAD) reveals keratin 8 as a novel candidate for interaction in pancreatic β-cells |
| title_short | Searching for protein partners of short-chain 3-hydroxyacyl-CoA dehydrogenase (SCHAD) reveals keratin 8 as a novel candidate for interaction in pancreatic β-cells |
| title_sort | searching for protein partners of short chain 3 hydroxyacyl coa dehydrogenase schad reveals keratin 8 as a novel candidate for interaction in pancreatic β cells |
| topic | Congenital hyperinsulinism of infancy Short-chain 3-hydroxyacyl-CoA dehydrogenase SCHAD HADH Intermediate filaments Keratin 8 |
| url | https://doi.org/10.1186/s12860-025-00544-w |
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