Pyruvate Oxidation !"#$%#&'(#(')*(+#(',-%&*(+#('.&%/*(#('_01'(%2$3(#+#('43&&*/5$0)%.43&&*/#('637#+#('8#$#('.&'(5$0)%.!"#$%&'(9.:;)%.04<))%.04<<=!<=!9.:9.:;)%.04<)%.04<>:?;?!*+!6.:9.:"#$%&'()*)+#*$,-).'/).@?;A)!=.:?;A?!"#$%&'()0'$1,2#3'/)<=!OCCCH3OOHPyruvateNAD+ CoA-SHNADH+CO2SCCH3OCoAAcetyl-CoA+ 3 NAD + FAD + GDP + PiTCA CyclePyruvateDehydrogenaseComplexCoA-SH + 3 NADH + FADH2 + GTP + 2 CO2Proteins in the Pyruvate Dehydrogenase Complex E1 = Pyruvate dehydrogenase (Pyruvate decarboxylase) E2 = Dihydrolipoyl acetyltransferase E3 = Dihydrolipoyl dehydrogenase Protein X E1-kinase Phospho-E1-phosphatase Size (megaDa) E1 peptides E2 peptides E3 peptides Eukaryotes gram-positive prokaryotes 10 120 ((αβ)2 tetramers) 60 (trimers) 24 (dimers) Gram-negative prokaryotes (E. coli) 4.6 24 24 12 Eukaryotic ribosome 4.2 4 RNA (6880 bases) and ~84 polypeptides Prokaryotic ribosome 2.7 3 RNA (4860 bases) and 57 polypeptides Thiamin pyrophosphateNNNSH3CNH2H3CHOPOOOPOOOHNSSOLipoamide•E2Lys-E2OCH2C CH3H3CCHHOCONHH2C CH2COHNOPOOOHOHCH2ONNNNNH2POOOCoenzyme A(CoA-SH)CH2CH2SHOOHOHCH2NOPOOOOOHOHCH2ONNNNNH2POONH2ONicotinamide adeninedinucleotide[NAD]OPOOOOOHOHCH2ONNNNNH2POONNNNHOOH3CH3COHOHHOFlavin adeninedinucleotide[FAD]Pyruvate Dehydrogenase Complex Thiamin pyrophosphateNNNSH3CNH2H3CHOPOOOPOOOHNSSOLys-E2(TPP•E1)HRNSRH3CHH3CCCOOOPyruvateRNSRH3CCH3COHCOOCO2RNSRH3CCH3COHLipoamide•E2TPP•E1HydroxyethylTPP•E1HNOLys-E2Acetyl-Lipoamide•E2SHSCOCH3CoA-SHCSOCH3CoAAcetyl-CoAHNOLys-E2Dihydrolipoamide•E2SHSHS SCysCysOxidized-E3[FAD]CysCysReduced-E3[FAD]SH SHTrimethylarsineArseniteClAsClClH3CAsCH3CH3OAsOOSHH2CHC SHH2C OHOAsRSHSHLipoamide+SSLipoamideAsROrganicArsenicalCu H AsO4NADNADHS SCysCysOxidized-E3[FAD]Tricarboxylic Acid (TCA) Cycle FADH2CO2H2CCOOOOOOHOH3C COS CoAOOOO OH2CCOOOOOOH2CCOOOOOOHOH2CCOOOOOOOH2CCOOOOOH2CCOOSOCoAOOOOOOOOOOO OOHCoA-SHH2OH2O1245678323GTPNADHNAD+CoA-SHCO2GDP + PiNADHFADH2GTPFADCoA-SH1. Citrate Synthase2. Aconitase3. Isocitrate Dehydrogenase4. !-Ketoglutarate Dehydrogenase5. Succinyl-CoA Synthetase6. Succinate Dehydrogenase7. Fumarase8. Malate DehydrogenaseNADNADHNADHNADNADHNADHCO2CO2H2OAcetyl-CoACitratecis-AconitateIsocitrate!-KetoglutarateOxalosuccinateSuccinyl-CoASuccinateFumarateMalateOxaloacetatePorcine (Sus scrofa) citrate synthase bound to citrate (pdb ID 1CTS) and to both citrate and CoA (pdb ID 2CTS) !""!#$!#"!""!""!#$"%&'(&)*+&+*,-)*+.'/!(-!0+1.'/!(-#$"!(-/2#!0+1&+*!"#$%#&'()*#+%,&-.%/0%1&#%#&!!"#$)/2!0331&#)/2!03!#$!"!(-23!""!#$!"!""44##056$7-5869:44##05$9;!""#!#$!"!(-23!""!#$!"!""44##056$7#-5869:44##05$9;!""#056$7<!(-=>(?@AB#056$7<C0+D(?+=!(-B!(-!0+1&+*E0+D(?+!(-E0+D!(-#05$9;!"#$%#&'()*#+%,&Reaction ∆G´° (kJ/mol) Typical ∆G in cells (kJ/mol) Acetate combustion -849 TCA cycle -40 -115 Citrate synthase -31.4 -53.9 Aconitase +6.7 +0.8 Isocitrate Dehydrogenase -8.4 -17.5 α-ketoglutarate dehydrogenase -30 -43.9 Succinyl-CoA synthetase -3.3 ~0 Succinate dehydrogenase +0.4 ~0 Fumarase -3.8 ~0 Malate dehydrogenase +29.7 ~0 !"!!""#"##!#$%&'()*+)#,-#./0*&1#2+%#$%%$&$&&%$%%$%%$&'()*+,-./-"01234-5678$&'$%%$%%$&'$%$%%$&$%%$%%$&'%3/4*)5++,6/-"$%_78"-*945-/./-"$&'$*1$%%$&'$%#5++,6:47$*1;#$&'$%%$%%$&'$%_78"-*945-/./-"_9:#)&1;<)+*+)#,-#./0*&1#2+%#$*17#&<012$%%$&'$%%$&&%$%%$&'$%$%%%3/4*/+"-/-"=/4/-"34-5=678>+;+)#,-#./0*&1#2+%#01234-5$%%$&'$&$%%$%%$&'$,-./-"4'&2()+%#%$%%$&$$%%$%%$&'!"!!""#"##!#&%&<&$%%$&$$%%$%%$&'!"!!""#"##!#&%&$%%$&$&&%$%%$%%$&'()*+,-./-"&<Anaplerotic reactions C H 3 C O C O O C H 2 C O C O O C O O C O 2 PyruvatePyruvatecarboxylaseATPADPPi++Oxaloacetate N H N S N H O O L y s - E n z y m e C O O O P O O O P O O O O H O H C H 2 O N N N N N H 2 P O O O O H C O O O P O O O C O O N H N S N H O O L y s - E n z y m e C H 2 C O C O O H O C O C H 2 C O C O O C O O C H 2 C O C O O ATPADPCarboxyphosphatePiCarboxybiotinylenzymeBiotinEnolpyruvatePyruvateOxaloacetate C H 2 C O O C H 2 C O C O O C H 2 C O O C H 2 C H C O O N H 3 + N H 4 + Glutamate!-ketoglutarateNADNADHGlutamatedehydrogenase O O O H O H C H 2 O N N N N N H 2 P O O N H 4 + O O O H O H C H 2 O N H N N N O P O O C H 2 C O O C H N H 3 + C O O C H 2 C O O C H N H C O O O O O H O H C H 2 O N N N N P O O A M P C H C O O C H C O O Adenosine Monophosphate(AMP)AMPdeaminaseInosine