Pyruvate dehydrogenase kinase (also pyruvate dehydrogenase complex kinase, PDC kinase, or PDK; EC 2.7.11.2) is a kinase enzyme which acts to inactivate the enzyme pyruvate dehydrogenase by phosphorylating it using ATP.
PDK thus participates in the regulation of the pyruvate dehydrogenase complex of which pyruvate dehydrogenase is the first component. Both PDK and the pyruvate dehydrogenase complex are located in the mitochondrial matrix of eukaryotes. The complex acts to convert pyruvate (a product of glycolysis in the cytosol) to acetyl-coA, which is then oxidized in the mitochondria to produce energy, in the citric acid cycle. By downregulating the activity of this complex, PDK will decrease the oxidation of pyruvate in mitochondria and increase the conversion of pyruvate to lactate in the cytosol.
The opposite action of PDK, namely the dephosphorylation and activation of pyruvate dehydrogenase, is catalyzed by a phosphoprotein phosphatase called pyruvate dehydrogenase phosphatase.
(Pyruvate dehydrogenase kinase should not be confused with Phosphoinositide-dependent kinase-1, which is also sometimes known as "PDK1".)
Phosphorylation Sites
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PDK can phosphorylate a serine residue on pyruvate dehydrogenase at three possible sites. Some evidence has shown that phosphorylation at site 1 is nearly completely deactivating while phosphorylation at sites 2 and 3 had only a small contribution to complex inactivation.[1] Therefore, it is phosphorylation at site 1 that is responsible for pyruvate dehydrogenase deactivation.
Isozymes
editThere are four known isozymes of PDK in humans:
The primary sequencing between the four isozymes are conserved with 70% identity. The greatest differences occur near the N-terminus.[2]
PDK1 is the largest of the four with 436 residues while PDK2, PDK3 and PDK4 have 407, 406, and 411 residues respectively. The isozymes have different activity and phosphorylation rates at each site. At site 1 in order from fastest to slowest, PDK2 > PDK4 ≈ PDK1 > PDK3. For site 2, PDK3 > PDK4 > PDK2 > PDK1. Only PDK1 can phosphorylate site 3. However, it has been shown that these activities are sensitive to slight changes in pH so the microenvironment of the PDK isozymes may change the reaction rates.[3][4]
Isozyme abundance has also been shown to be tissue specific. PDK1 is ample in heart cells. PDK3 is most abundant in testis. PDK2 is present in most tissues but low in spleen and lung cells. PDK4 is predominantly found in skeletal muscle and heart tissues. [5]
Mechanism
editPyruvate dehydrogenase is deactivated when phosphorylated by PDK. Normally, the active site of pyruvate dehydrogenase is stabilized into an ordered conformation through a hydrogen bond network. However, phosphorylation by PDK at site 1 causes steric clashes with another nearby serine residue due to both the increased size of the residue and negative charges.[6] This disrupts the hydrogen bond network and disorders the conformation of two phosphorylation loops. These loops prevent the reductive acetylation step, thus halting overall activity of the enzyme.[7] The conformational changes and mechanism of deactivation for phosphorylation at sites 2 and 3 are not known at this time.
Regulation
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Pyruvate dehydrogenase kinase is stimulated by ATP, NADH and acetyl-CoA. It is inhibited by ADP, NAD+, CoA-SH and pyruvate.[9]
Each isozyme responds slightly differently to each of these factors. NADH stimulates PDK1 activity by 20% and PDK2 activity by 30%. NADH with acetyl-CoA increases activity in these enzymes by 200 and 300% respectively. In similar conditions, PDK3 is unresponsive to NADH and inhibited by NADH with acetyl-CoA. PDK4 has an activity increase of 200% with NADH, but adding acetyl-CoA does not increase activity further.[5]
Disease Relevance
editSome studies have shown that in cells that lack insulin (or are insensitive to insulin) PDK4 is overexpressed.[10] As a result, the pyruvate formed from glycolysis cannot be oxidized which will lead to hyperglycaemia since the glucose in the blood cannot be efficiently used. Therefore several drugs target PDK4 hoping to treat type II diabetes.[11]
PDK1 has shown to have increased activity in hypoxic cancer cells due to the presence of HIF-1. PDK1 shunts pyruvate away from the citric acid cycle and keeps the hypoxic cell alive.[12] Therefore, PDK1 inhibition has been suggested as an antitumor therapy since PDK1 prevents apoptosis in these cancerous cells.[13] Similarly, PDK3 has been shown to be overexpressed in colon cancer cell lines.[14] Three proposed inhibitors are AZD7545 and Dichloroacetate which both bind to PDK1, and Radicicol which binds to PDK3.[15]
References
edit- ^ Yeaman, Stephen J., David C. Watson, and Gordon H. Dixon. "Sites of phosphorylation on pyruvate dehydrogenase from bovine kidney and heart." Biochemistry Vol. 17. Issue 12 (1978): 2364-369. http://pubs.acs.org/doi/pdf/10.1021/bi00605a017
- ^ Popov KM, Kedishvili NY, Zhao Y, Gudi R and Harris RA. “Molecular Cloning of the p45 subunit of pyruvate dehydrogenase kinase.” J. Biol. Chem., Vol. 269, Issue 47, 29720-29724, Nov, 1994, http://hwmaint.jbc.org/cgi/content/abstract/269/47/29720
- ^ Korotchkina L and Patel M. “Site specificity of four pyruvate dehydrogenase kinase isoenzymes toward the three phosphorylation sites of human pyruvate dehydrogenase.” J. Biol. Chem., Vol. 276, Issue 40: 37223-37229, Oct. 2001. http://www.jbc.org/content/276/40/37223.full.pdf+html
- ^ Kolobova E, Tuganova A, Boulatnikov I, and Popov K. “Regulation of pyruvate dehydrogenase activity through phosphorylation at multiple sites.” Biochem. J. Vol. 358: 69-77, Aug, 2001. http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1222033/
- ^ a b Bowker-Kinley MM, Davis WI, Wu P, Harris RA, and Popov KM. “Evidence for the existence of tissue-specific regulation of the mammalian pyruvate dehydrogenase complex.” Biochem. J. Vol. 329: 191-196, Jan. 1998. http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1219031/
- ^ Korotchkina LG and Patel MS. “Probing the mechanism of inactivation of human pyruvate dehydrogenase by phosphorylation of three sites.” J. Biol. Chem. Vol. 276, No. 8: 5731-5738, Feb. 2001. http://hwmaint.jbc.org/cgi/reprint/276/8/5731
- ^ Kato M, Wynn RM, Chuang JL, Tso SC, Machius M, Li J, and Chuang DT. “Structural basis for inactivation of human pyruvate dehydrogenase complex by phosphorylation: role of disordered phosphorylation loops.” Structure. Vol. 16, Issue 12: 1849-1859, Dec. 2008. http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2849990/
- ^ Roche TE and Reed LJ. “Monovalent cation requirement for ADP inhibition of pyruvate dehydrogenase kinase.” Biochemical and Biophysical Research Communications. Vol. 59, Issue 4: 1341-1348, Aug. 1974.
- ^ Roche TE and Reed LJ. “Monovalent cation requirement for ADP inhibition of pyruvate dehydrogenase kinase.” Biochemical and Biophysical Research Communications. Vol. 59, Issue 4: 1341-1348, Aug. 1974. http://www.sciencedirect.com/science/article/pii/0006291X74904616
- ^ Majer M, Popov KM, Harris RA, Bogardus C and Prochazka M. “Insulin downregulates pyruvate dehydrogenase kinase mRNA: potential mechanism contributing to increased lipid oxidation in insulin-resistant subjects.” Molecular Genetics and Metabolism. Vol. 65, Issue 2: 181-186, Oct. 1998. http://www.sciencedirect.com/science/article/pii/S1096719298927482
- ^ Holness MJ and Sugden MC. “Regulation of pyruvate dehydrogenase complex activity by reversible phosphorylation.” Biochemical Society Transactions. Vol. 31, pt. 6: 1143-1151. 2003. http://www.ncbi.nlm.nih.gov/pubmed/14641014
- ^ Kim J, Tchernyshyov I, Semenza G and Dang CV. “HIF-1-mediated expression of pyruvate dehydrogenase kinase: a metabolic switch required for cellular adaptation to hypoxia.” Cell Metabolism. Vol. 3, Issue 3: 177-185. March 2006. http://www.sciencedirect.com/science/article/pii/S1550413106000623
- ^ Bonnet S, Michelakis ED, et. al. “A mitochondria-K+ channel axis is suppressed in cancer and its normalization promotes apoptosis and inhibits cancer growth.” Cancer Cell. Vol. 11, Issue 1: 37-51. Jan. 2007. http://www.sciencedirect.com/science/article/pii/S1535610806003722
- ^ Lu CW, Lin CS, Chein CW, Lin SC, Lee CT, Lin BW, Lee JC and Tsai SJ. “Overexpression of pyruvate dehydrogenase kinase 3 increases drug resistance and early recurrence in colon cancer.” The American Journal of Pathology. Vol. 179, Issue 3: 1405-1414. Sept. 2011. http://www.sciencedirect.com/science/article/pii/S0002944011005499
- ^ Kato M, Li J, Chuang JL and Chuang DT. “Distinct structural mechanisms for inhibition of pyruvate dehydrogenase kinase isoforms by AZD7545, dichloroacetate, and radicicol.” Structure. Vol. 15, Issue 8: 992-1004. Aug 2007. http://www.sciencedirect.com/science/article/pii/S096921260700250X
External links
edit- pyruvate+dehydrogenase+kinase at the U.S. National Library of Medicine Medical Subject Headings (MeSH)
- EC 2.7.11.2
Category:EC 2.7.11 Category:Citric acid cycle Category:Glycolysis