The mature protein encoded by thePDK4 gene contains 294 amino acids in its sequence. To form the active protein, two of the polypeptide chains come together to form an open conformation.[6] Specifically, the two subunits come together to form a nucleotide-binding pocket; this pocket is targeted most often by inhibitors.[8]
The pyruvate dehydrogenase (PDH) complex must be tightly regulated due to its central role in general metabolism. Within the complex, there are three serine residues on the E1 component that are sites for phosphorylation; this phosphorylation inactivates the complex. In humans, there have been fourisozymes of pyruvate dehydrogenase kinase that have been shown to phosphorylate these three sites:PDK1,PDK2,PDK3, and PDK4. PDK4 does not incorporate the most phosphate groups per catalytic event, because it can only phosphorylate site 1 and site 2; its rate of phosphorylation is less than PDK1, equal to PDK3, and more than PDK2. When thethiamine pyrophosphate (TPP)coenzyme is bound, the rates of phosphorylation by all four isozymes are drastically affected. Site 1 is the most affected, with the rate being significantly decreased. However, overall activity by PDK4 is not affected.[9]
As the primary regulators of a crucial step in the central metabolic pathway, the pyruvate dehydrogenase family is tightly regulated itself by a myriad of factors including transcription factorsSp1 andCCAAT box binding factor (CBF).Retinoic acid enhancesPDK4 transcription by enablingretinoic acid receptor family members to recruit transcriptional coactivators to retinoic acid response elements (RAREs) in thePDK4 promoter. Transcription is also increased by inhibiting inhibitoryhistone deacetylases (HDACs) usingtrichostatin A (TSA).[10]Rosiglitazone, athiazolidinedione known to activate theglycerol biogenesis pathway, increasesPDK4mRNA transcription inwhite adipose tissue, but not in liver or muscle tissue.[11]Farnesoid X receptor, or FXR, suppresses glycolysis and enhancesfatty acid oxidation by increasingPDK4 expression and inactivating the PDH complex.[12]Other factors, such asinsulin, directly downregulate bothPDK2 andPDK4 mRNA transcription. This is done through a proposedphosphatidylinositol 3-kinase (PI3K)-dependent pathway. In fact, even when cells are exposed todexamethasone to increase mRNA expression, insulin blocks this effect.[13]Peroxisome proliferator-activated receptors also regulate expression; PPAR alpha and delta were found to upregulatePDK4 mRNA, but PPAR gamma activation downregulated expression.[14]
PDK4 is relevant in a variety of clinical conditions. Short-term fasting induces an increase inPDK4 transcription by about 10-fold.[15] Upon refeeding, transcription of PDK4 increased further, a surprising outlook, by about 50-fold over levels before fasting began.[16] This effect can be seen long term as well. PDK4 is overexpressed in skeletal muscle intype 2 diabetes, resulting in impaired glucose utilization.[17] In post-obese patients, there is a significant decrease inPDK4 mRNA expression, in conjunction with increased glucose uptake; this is likely due to the downregulation of PDK4 by insulin. This corroborates the concept that a lowered availability of free fatty acids affects glucose metabolism by PDH complex regulation.[18]In fact, it has been shown that insufficient downregulation of PDK mRNA in insulin-resistant individuals could be a cause of increased PDK expression leading to impaired glucose oxidation followed by increased fatty acid oxidation.[19]
Exercise has been shown to induce changes in this gene as well, and that transient changes can have a cumulative effect across many exercise sessions. The mRNA ofPDK4, along withPPARGC1A, increases in both types of muscle tissue after exercise.[20][21]
These metabolic effects can be seen in other conditions.Hypoxia is shown to induce PDK4 gene expression through theERR gamma mechanism.[22]Conversely,PDK4 is downregulated in cardiac muscle tissue during heart failure.[23]
The ubiquitous role of this gene lends itself to being involved in a variety of disease pathologies, including cancer. One metabolite,butyrate, induceshyperacetylation of the histones around thePDK4 gene. This is associated with a greater transcription level ofPDK4 mRNA, thereby reversing the downregulation of PDK4 in colon carcinoma cells. In human colon cancer cells, targeting and inactivating the PDH complex limits the metabolic rate and regulatesglutamine metabolism, thereby partially inhibiting cell growth.[24] However, PDK4 has also been shown to promotetumorigenesis and proliferation through a different pathway, theCREB-RHEB-mTORC1 signaling cascade.[25]
^Kukimoto-Niino M, Tokmakov A, Terada T, Ohbayashi N, Fujimoto T, Gomi S, Shiromizu I, Kawamoto M, Matsusue T, Shirouzu M, Yokoyama S (Sep 2011). "Inhibitor-bound structures of human pyruvate dehydrogenase kinase 4".Acta Crystallographica Section D.67 (Pt 9):763–73.doi:10.1107/S090744491102405X.PMID21904029.
^Kwon HS, Huang B, Ho Jeoung N, Wu P, Steussy CN, Harris RA (2006). "Retinoic acids and trichostatin A (TSA), a histone deacetylase inhibitor, induce human pyruvate dehydrogenase kinase 4 (PDK4) gene expression".Biochimica et Biophysica Acta (BBA) - Gene Structure and Expression.1759 (3–4):141–51.doi:10.1016/j.bbaexp.2006.04.005.PMID16757381.
^Savkur RS, Bramlett KS, Michael LF, Burris TP (Apr 2005). "Regulation of pyruvate dehydrogenase kinase expression by the farnesoid X receptor".Biochemical and Biophysical Research Communications.329 (1):391–6.doi:10.1016/j.bbrc.2005.01.141.PMID15721319.
^Spriet LL, Tunstall RJ, Watt MJ, Mehan KA, Hargreaves M, Cameron-Smith D (Jun 2004). "Pyruvate dehydrogenase activation and kinase expression in human skeletal muscle during fasting".Journal of Applied Physiology.96 (6):2082–7.doi:10.1152/japplphysiol.01318.2003.PMID14966024.S2CID13601849.
^Pilegaard H, Ordway GA, Saltin B, Neufer PD (Oct 2000). "Transcriptional regulation of gene expression in human skeletal muscle during recovery from exercise".American Journal of Physiology. Endocrinology and Metabolism.279 (4): E806-14.doi:10.1152/ajpendo.2000.279.4.e806.PMID11001762.S2CID1008940.
^Wang L, Sahlin K (Apr 2012). "The effect of continuous and interval exercise on PGC-1α and PDK4 mRNA in type I and type II fibres of human skeletal muscle".Acta Physiologica.204 (4):525–32.doi:10.1111/j.1748-1716.2011.02354.x.PMID21883960.S2CID13208033.
^Razeghi P, Young ME, Ying J, Depre C, Uray IP, Kolesar J, Shipley GL, Moravec CS, Davies PJ, Frazier OH, Taegtmeyer H (2002). "Downregulation of metabolic gene expression in failing human heart before and after mechanical unloading".Cardiology.97 (4):203–9.doi:10.1159/000063122.PMID12145475.S2CID46861699.
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Razeghi P, Young ME, Ying J, Depre C, Uray IP, Kolesar J, Shipley GL, Moravec CS, Davies PJ, Frazier OH, Taegtmeyer H (2002). "Downregulation of metabolic gene expression in failing human heart before and after mechanical unloading".Cardiology.97 (4):203–9.doi:10.1159/000063122.PMID12145475.S2CID46861699.
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