Cardiac ATP-sensitive K+ channel associates with the glycolytic enzyme complex
| Autor: | Brian Malester, Miyoun Hong, Eirini Kefaloyianni, Li Bao, Thomas A. Neubert, Diane Delaroche, William A. Coetzee |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2011 |
| Předmět: |
Male
Enzyme complex endocrine system Receptors Drug Immunoblotting Fructose-bisphosphate aldolase Sulfonylurea Receptors Biochemistry Research Communications Rats Sprague-Dawley KATP Channels Tandem Mass Spectrometry Fructose-Bisphosphate Aldolase Two-Hybrid System Techniques Chlorocebus aethiops Genetics Animals Humans Immunoprecipitation Glycolysis Potassium Channels Inwardly Rectifying Molecular Biology Gene Library chemistry.chemical_classification biology Myocardium Aldolase A Cell Membrane Kir6.2 Rats Enzyme HEK293 Cells chemistry COS Cells biology.protein Sulfonylurea receptor ATP-Binding Cassette Transporters Glyceraldehyde-3-Phosphate Dehydrogenase (Phosphorylating) Pyruvate kinase Biotechnology Protein Binding |
| Popis: | Being gated by high-energy nucleotides, cardiac ATP-sensitive potassium (KATP) channels are exquisitely sensitive to changes in cellular energy metabolism. An emerging view is that proteins associated with the KATP channel provide an additional layer of regulation. Using putative sulfonylurea receptor (SUR) coiled-coil domains as baits in a 2-hybrid screen against a rat cardiac cDNA library, we identified glycolytic enzymes (GAPDH and aldolase A) as putative interacting proteins. Interaction between aldolase and SUR was confirmed using GST pulldown assays and coimmunoprecipitation assays. Mass spectrometry of proteins from KATP channel immunoprecipitates of rat cardiac membranes identified glycolysis as the most enriched biological process. Coimmunoprecipitation assays confirmed interaction for several glycolytic enzymes throughout the glycolytic pathway. Immunocytochemistry colocalized many of these enzymes with KATP channel subunits in rat cardiac myocytes. The catalytic activities of aldolase and pyruvate kinase functionally modulate KATP channels in patch-clamp experiments, whereas d-glucose was without effect. Overall, our data demonstrate close physical association and functional interaction of the glycolytic process (particularly the distal ATP-generating steps) with cardiac KATP channels.—Hong, M., Kefaloyianni, E., Bao, L., Malester, B., Delaroche, D., Neubert, T. A., Coetzee, W. A. Cardiac ATP-sensitive K+ channel associates with the glycolytic enzyme complex. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|