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The glycolytic enzymes, glyceraldehyde-3-phosphate dehydrogenase, triose-phosphate isomerase, and pyruvate kinase are components of the K(ATP) channel macromolecular complex and regulate its function.
J Biol Chem. 2005 Nov 18; 280(46):38464-70.JB

Abstract

The regulation of ATP-sensitive potassium (K(ATP)) channel activity is complex and a multitude of factors determine their open probability. Physiologically and pathophysiologically, the most important of these are intracellular nucleotides, with a long-recognized role for glycolytically derived ATP in regulating channel activity. To identify novel regulatory subunits of the K(ATP) channel complex, we performed a two-hybrid protein-protein interaction screen, using as bait the mouse Kir6.2 C terminus. Screening a rat heart cDNA library, we identified two potential interacting proteins to be the glycolytic enzymes, glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and triose-phosphate isomerase. The veracity of interaction was verified by co-immunoprecipitation techniques in transfected mammalian cells. We additionally demonstrated that pyruvate kinase also interacts with Kir6.2 subunits. The physiological relevance of these interactions is illustrated by the demonstration that native Kir6.2 protein similarly interact with GAPDH and pyruvate kinase in rat heart membrane fractions and that Kir6.2 protein co-localize with these glycolytic enzymes in rat ventricular myocytes. The functional relevance of our findings is demonstrated by the ability of GAPDH or pyruvate kinase substrates to directly block the K(ATP) channel under patch clamp recording conditions. Taken together, our data provide direct evidence for the concept that key enzymes involved in glycolytic ATP production are part of a multisubunit K(ATP) channel protein complex. Our data are consistent with the concept that the activity of these enzymes (possibly by ATP formation in the immediate intracellular microenvironment of this macromolecular K(ATP) channel complex) causes channel closure.

Authors+Show Affiliations

Department of Pediatrics, New York University School of Medicine, New York, New York 10016, USA.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

16170200

Citation

Dhar-Chowdhury, Piyali, et al. "The Glycolytic Enzymes, Glyceraldehyde-3-phosphate Dehydrogenase, Triose-phosphate Isomerase, and Pyruvate Kinase Are Components of the K(ATP) Channel Macromolecular Complex and Regulate Its Function." The Journal of Biological Chemistry, vol. 280, no. 46, 2005, pp. 38464-70.
Dhar-Chowdhury P, Harrell MD, Han SY, et al. The glycolytic enzymes, glyceraldehyde-3-phosphate dehydrogenase, triose-phosphate isomerase, and pyruvate kinase are components of the K(ATP) channel macromolecular complex and regulate its function. J Biol Chem. 2005;280(46):38464-70.
Dhar-Chowdhury, P., Harrell, M. D., Han, S. Y., Jankowska, D., Parachuru, L., Morrissey, A., Srivastava, S., Liu, W., Malester, B., Yoshida, H., & Coetzee, W. A. (2005). The glycolytic enzymes, glyceraldehyde-3-phosphate dehydrogenase, triose-phosphate isomerase, and pyruvate kinase are components of the K(ATP) channel macromolecular complex and regulate its function. The Journal of Biological Chemistry, 280(46), 38464-70.
Dhar-Chowdhury P, et al. The Glycolytic Enzymes, Glyceraldehyde-3-phosphate Dehydrogenase, Triose-phosphate Isomerase, and Pyruvate Kinase Are Components of the K(ATP) Channel Macromolecular Complex and Regulate Its Function. J Biol Chem. 2005 Nov 18;280(46):38464-70. PubMed PMID: 16170200.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - The glycolytic enzymes, glyceraldehyde-3-phosphate dehydrogenase, triose-phosphate isomerase, and pyruvate kinase are components of the K(ATP) channel macromolecular complex and regulate its function. AU - Dhar-Chowdhury,Piyali, AU - Harrell,Maddison D, AU - Han,Sandra Y, AU - Jankowska,Danuta, AU - Parachuru,Lavanya, AU - Morrissey,Alison, AU - Srivastava,Shekhar, AU - Liu,Weixia, AU - Malester,Brian, AU - Yoshida,Hidetada, AU - Coetzee,William A, Y1 - 2005/09/16/ PY - 2005/9/20/pubmed PY - 2006/1/13/medline PY - 2005/9/20/entrez SP - 38464 EP - 70 JF - The Journal of biological chemistry JO - J. Biol. Chem. VL - 280 IS - 46 N2 - The regulation of ATP-sensitive potassium (K(ATP)) channel activity is complex and a multitude of factors determine their open probability. Physiologically and pathophysiologically, the most important of these are intracellular nucleotides, with a long-recognized role for glycolytically derived ATP in regulating channel activity. To identify novel regulatory subunits of the K(ATP) channel complex, we performed a two-hybrid protein-protein interaction screen, using as bait the mouse Kir6.2 C terminus. Screening a rat heart cDNA library, we identified two potential interacting proteins to be the glycolytic enzymes, glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and triose-phosphate isomerase. The veracity of interaction was verified by co-immunoprecipitation techniques in transfected mammalian cells. We additionally demonstrated that pyruvate kinase also interacts with Kir6.2 subunits. The physiological relevance of these interactions is illustrated by the demonstration that native Kir6.2 protein similarly interact with GAPDH and pyruvate kinase in rat heart membrane fractions and that Kir6.2 protein co-localize with these glycolytic enzymes in rat ventricular myocytes. The functional relevance of our findings is demonstrated by the ability of GAPDH or pyruvate kinase substrates to directly block the K(ATP) channel under patch clamp recording conditions. Taken together, our data provide direct evidence for the concept that key enzymes involved in glycolytic ATP production are part of a multisubunit K(ATP) channel protein complex. Our data are consistent with the concept that the activity of these enzymes (possibly by ATP formation in the immediate intracellular microenvironment of this macromolecular K(ATP) channel complex) causes channel closure. SN - 0021-9258 UR - https://www.unboundmedicine.com/medline/citation/16170200/The_glycolytic_enzymes_glyceraldehyde_3_phosphate_dehydrogenase_triose_phosphate_isomerase_and_pyruvate_kinase_are_components_of_the_K_ATP__channel_macromolecular_complex_and_regulate_its_function_ L2 - http://www.jbc.org/cgi/pmidlookup?view=long&pmid=16170200 DB - PRIME DP - Unbound Medicine ER -