Regulation of mammalian pyruvate dehydrogenase complex by phosphorylation: complexity of multiple phosphorylation sites and kinases

This review summarizes the recent developments on the regulation of human pyruvate dehydrogenase complex (PDC) by site-specific phosphorylation by four kinases. Mutagenic analysis of the three phosphorylation sites of human pyruvate dehydrogenase (E1) showed the site-independent mechanism of phosphorylation as well as site-independent dephosphorylation of the three phosphorylation sites and the importance of each phosphorylation site for the inactivation of E1. Both the negative charge and size of the group introduced at site 1 were involved in human E1 inactivation. Mechanism of inactivation of E1 was suggested to be site-specific. Phosphorylation of site 1 affected E1 interaction with the lipoyl domain of dihydrolipoamide acetyltransferase, whereas phosphorylation site 3 appeared to be closer to the thiamine pyrophosphate (TPP)-binding region affecting coenzyme interaction with human E1. Four isoenzymes of pyruvate dehydrogenase kinase (PDK) showed different specificity for the three phosphorylation sites of E1. All four PDKs phosphorylated sites 1 and 2 in PDC with different rates, and only PDK1 phosphorylated site 3. PDK2 was maximally stimulated by the reduction/acetylation of the lipoyl groups of E2. Presence of the multiple phosphorylation sites and isoenzymes of PDK is important for the tissue-specific regulation of PDC under different physiological conditions.

[1]  G. Brown,et al.  The human pyruvate dehydrogenase complex. Isolation of cDNA clones for the E1 alpha subunit, sequence analysis, and characterization of the mRNA. , 1987, The Journal of biological chemistry.

[2]  Randall,et al.  Partial purification and characterization of the maize mitochondrial pyruvate dehydrogenase complex , 1998, Plant physiology.

[3]  L. Reed,et al.  A Trail of Research from Lipoic Acid to α-Keto Acid Dehydrogenase Complexes , 2001, The Journal of Biological Chemistry.

[4]  R. Harris,et al.  Expression and regulation of pyruvate dehydrogenase kinase isoforms in the developing rat heart and in adulthood: role of thyroid hormone status and lipid supply. , 2000, The Biochemical journal.

[5]  T. Roche,et al.  Function of the nonidentical subunits of mammalian pyruvate dehydrogenase. , 1972, Biochemical and biophysical research communications.

[6]  J. R. Butler,et al.  Binding of thiamin thiazolone pyrophosphate to mammalian pyruvate dehydrogenase and its effects of kinase and phosphatase activities. , 1977, Biochemical and biophysical research communications.

[7]  T. Roche,et al.  Requirements for the Adaptor Protein Role of Dihydrolipoyl Acetyltransferase in the Up-regulated Function of the Pyruvate Dehydrogenase Kinase and Pyruvate Dehydrogenase Phosphatase* , 1998, The Journal of Biological Chemistry.

[8]  M. Holness,et al.  Interactive regulation of the pyruvate dehydrogenase complex and the carnitine palmitoyltransferase system , 1994, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[9]  K. M. Popov,et al.  Evidence for existence of tissue-specific regulation of the mammalian pyruvate dehydrogenase complex. , 1998, The Biochemical journal.

[10]  K. M. Popov,et al.  Site-directed mutagenesis of phosphorylation sites of the branched chain alpha-ketoacid dehydrogenase complex. , 1994, The Journal of biological chemistry.

[11]  Y. J. Huang,et al.  Expression of pyruvate dehydrogenase isoforms during the aerobic/anaerobic transition in the development of the parasitic nematode Ascaris suum: altered stoichiometry of phosphorylation/inactivation. , 1998, Archives of biochemistry and biophysics.

[12]  S. Galadari,et al.  Effects of cations on ceramide-activated protein phosphatase 2A , 2001, Experimental & Molecular Medicine.

[13]  G. J. Sale,et al.  Occupancy of phosphorylation sites in pyruvate dehydrogenase phosphate complex in rat heart in vivo. Relation to proportion of inactive complex and rate of re-activation by phosphatase. , 1982, The Biochemical journal.

[14]  Hyung-Lae Kim,et al.  Cleavage of purified neuronal clathrin assembly protein (CALM) by caspase 3 and calpain , 2001, Experimental & Molecular Medicine.

[15]  J. Thelen,et al.  Pyruvate dehydrogenase kinase from Arabidopsis thaliana: a protein histidine kinase that phosphorylates serine residues. , 2000, The Biochemical journal.

[16]  P. Komuniecki,et al.  Molecular cloning, functional expression, and characterization of pyruvate dehydrogenase kinase from anaerobic muscle of the parasitic nematode Ascaris suum. , 1998, Archives of biochemistry and biophysics.

[17]  W. Hol,et al.  Crystal structure of human branched-chain α-ketoacid dehydrogenase and the molecular basis of multienzyme complex deficiency in maple syrup urine disease , 2000 .

[18]  R. Harris,et al.  Control of pyruvate dehydrogenase kinase gene expression. , 2001, Advances in enzyme regulation.

[19]  W. Hol,et al.  Crystal structure of 2-oxoisovalerate and dehydrogenase and the architecture of 2-oxo acid dehydrogenase multienzyme complexes , 1999, Nature Structural Biology.

[20]  M. Patel,et al.  Site Specificity of Four Pyruvate Dehydrogenase Kinase Isoenzymes toward the Three Phosphorylation Sites of Human Pyruvate Dehydrogenase* , 2001, The Journal of Biological Chemistry.

[21]  Jianchun Dong,et al.  Distinct regulatory properties of pyruvate dehydrogenase kinase and phosphatase isoforms. , 2001, Progress in nucleic acid research and molecular biology.

[22]  T. Roche,et al.  Lipoyl Domain-based Mechanism for the Integrated Feedback Control of the Pyruvate Dehydrogenase Complex by Enhancement of Pyruvate Dehydrogenase Kinase Activity (*) , 1996, The Journal of Biological Chemistry.

[23]  T. Roche,et al.  Marked Differences between Two Isoforms of Human Pyruvate Dehydrogenase Kinase* , 2000, The Journal of Biological Chemistry.

[24]  M. Patel,et al.  Mutagenesis Studies of the Phosphorylation Sites of Recombinant Human Pyruvate Dehydrogenase. SITE-SPECIFIC REGULATION (*) , 1995, The Journal of Biological Chemistry.

[25]  D. Walsh,et al.  The elementary reactions of the pig heart pyruvate dehydrogenase complex. A study of the inhibition by phosphorylation. , 1976, The Biochemical journal.

[26]  T. Roche,et al.  Molecular biology and biochemistry of pyruvate dehydrogenase complexes 1 , 1990, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[27]  L. Korotchkina,et al.  Probing the Mechanism of Inactivation of Human Pyruvate Dehydrogenase by Phosphorylation of Three Sites* , 2001, The Journal of Biological Chemistry.

[28]  R. Perham,et al.  Swinging arms and swinging domains in multifunctional enzymes: catalytic machines for multistep reactions. , 2000, Annual review of biochemistry.

[29]  R. Harris,et al.  Mammalian α‐keto acid dehydrogenase complexes: gene regulation and genetic defects 1 , 1995, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[30]  K. M. Popov,et al.  Evidence that pyruvate dehydrogenase kinase belongs to the ATPase/kinase superfamily. , 1999, The Biochemical journal.

[31]  L. S. Khailova,et al.  Chemical modification of the essential arginine residues of pyruvate dehydrogenase prevents its phosphorylation by kinase , 1996, FEBS letters.

[32]  K. M. Popov,et al.  Regulation of pyruvate dehydrogenase activity through phosphorylation at multiple sites. , 2001, The Biochemical journal.

[33]  K. M. Popov,et al.  Starvation and diabetes increase the amount of pyruvate dehydrogenase kinase isoenzyme 4 in rat heart. , 1998, The Biochemical journal.

[34]  J. R. Brown,et al.  Sites of phosphorylation on pyruvate dehydrogenase from bovine kidney and heart. , 1978, Biochemistry.

[35]  K. M. Popov,et al.  Cloning and Characterization of PDK4 on 7q21.3 Encoding a Fourth Pyruvate Dehydrogenase Kinase Isoenzyme in Human* , 1996, The Journal of Biological Chemistry.

[36]  T. Roche,et al.  Reaction mechanism for mammalian pyruvate dehydrogenase using natural lipoyl domain substrates. , 2001, Archives of biochemistry and biophysics.

[37]  K. M. Popov,et al.  Starvation increases the amount of pyruvate dehydrogenase kinase in several mammalian tissues. , 2000, Archives of biochemistry and biophysics.

[38]  P. Sugden,et al.  Regulation of pig heart pyruvate dehydrogenase by phosphorylation. Studies on the subunit and phosphorylation stoicheiometries. , 1978, The Biochemical journal.

[39]  R. Ueda,et al.  cDNA sequence and expression of a gene encoding a pyruvate dehydrogenase kinase homolog of Drosophila melanogaster. , 1997, DNA and cell biology.

[40]  L. Korotchkina,et al.  The effect of phosphorylation on pyruvate dehydrogenase , 1995, FEBS letters.