Acetyl-CoA + FAD + GDP + Pi + 2H2O + 3NAD+ -> CoA + 2CO2 + FADH2 + GTP + 3NADH + 3H+ ; This reaction takes place in the mitrochondrial matrix.
Write the overall equation for the TCA Cycle. Where does it take place?
acetyl CoA C2 -> citrate C6 -> cis-aconitase C6 -> isocitrate C6 -> a-ketoglutarate C5 -> succinyl CoA C4 -> succinate -> C4 -> fumarate C4 -> malate C4 -> oxeloacetate C4
Starting with acetyl-CoA (C2), what is the number of carbons at each step of the cycle?
a-ketoglutarate dehydrogenase complex
What enzyme catalyzes the conversion of a-ketoglutarate to succinyl CoA?
a-ketoglutarate -> succinyl CoA
What reaction does the a-ketoglutarate dehydrogenase complex catalyze?
isocitrate -> a-ketoglutarate and a-ketoglutarate -> succinyl CoA
Which step(s) in the TCA cycle involve the formation of CO2 (ie. The decarboxylation steps)?
isocitrate -> a-ketoglutarate; a-ketoglutarate -> succinyl CoA; malate -> oxaloacetate
Which step(s) in the TCA cycle involve the formation of NADH + H+?
Succinyl-CoA + GDP + Pi → Succinate + GTP → ATP
Which step(s) in the TCA cycle involve the formation of ATP (by substrate level phosphorylation?)
10 ATP are liberated from one turn of the cycle.
The total number of ATP that can be generated from one turn of the cycle (include any from oxidative phosphorylation)?
Isocitrate dehydrogenase and a-ketoglutarate dehydrogenase are inhibited by NADH and ATP.
What enzymes are involved in regulating the TCA cycle?
Isocitrate dehydrogenase is stimulated by ADP, and inhibited by both ATP and NADH. A-KG is inhibited by succinyl CoA, NADH, and ATP.
How are isocitrate dehydrogenase and a-ketoglutarate dehydrogenase turned on and off?
1) Pyruvate + CoA + NAD+ → AcCoA + NADH + H+ + CO2 and 2)αKG + CoA + NAD+ → SuccCoA + NADH + H+ + CO2. The reactions proceed in an analogous manner and require the same five coenzymes (TPP, Lipoic Acid, CoA, FAD, and NAD+).
What are the similarities between the oxidative decarboxylation of pyruvate and a-ketoglutarate?
TPP (Thiamine Pyrophosphate), Lipoic Acid, CoA, FAD, NAD+
Name the 5 coenzymes which participate in the pyruvate → acetyl CoA and α-Ketoglutarate → succinyl CoA steps.
This term is used to describe reactions which form intermediates of the TCA cycle.
There are a few anaplerotic reactions, but the one of most physiological importance is Pyruvate -> Oxaloacetate via the enzyme pyruvate carboxylase.
Which intermediate of the TCA cycle can be replenished by an anaplerotic reaction?
Pyruvate carboxylase catalyzes the reaction and is activated by acetyl-CoA; biotin is the coenzyme that is required in order for pyruvate to be carboxylated.
What enzyme catalyzes Pyruvate -> Oxaloacetate? What regulates this enzyme? What coenzyme is required?
Glutamate can be converted to a-KG via glutamate dehydrogenase.
How can a-ketoglutarate be regenerated? Include precursor and enzyme.
Biotin is a coenzyme which facilitates the carboxylation of pyruvate. Biotin provides an active form of CO2.
What is the role of biotin in the conversion of pyruvate->oxaloacetate?
The reaction takes place in the mitochondrial matrix. It is important for the metabolism of glucose because it is irreversible and the reaction must proceed into the TCA cycle. Otherwise, pyruvate will be converted to lactic acid.
Where does the following reaction take place and why is it so important in the metabolism of glucose? Pyruvate->AcetylCoA
There is a symport protein carrier that allows pyruvate to cross into the mitochondrial matrix.
How does pyruvate traverse the inner mitochondrial membrane?
Ca+2 activates PDH phosphatase, which dephosphorylates PDH and makes it active. ATP is a substrate for PDH kinase, which phosphorylizes PDH and makes it inactive. PDH is also regulated by product inhibition through acetyl-CoA and NADH. Finally PDH is regulated by the need for adequate amounts of NAD+ and CoA.
How is pyruvate dehydrogenase regulated?
Thiamine is a co-factor for transketolase (in pentose phosphate pathway) and a coenzyme for the decarboxylation of a-keto acids.
In general, what type of reaction(s) does thiamine (Vitamin B1) participate as a co-enzyme?
Beri Beri (Dry or Wet), and Wernicke-Korsakoff syndrome
What diseases results from a deficiency of thiamine?
Measure the ratio of lactic acid to pyruvic acid in the blood after administration of glucose or measure the amount of transketolase present in RBCs. Blood and urinary levels of pyruvic acid are characteristically elevated in thiamine deficiency but the ratio is a better indicator.
Describe a diagnostic test for thiamine deficiency?
The carboxyl group in lipoic acid is important because it binds covalently with amino-Lysine of the enzyme (E2). Forms an R-C=O-NH-Lys-Enz. (Amide Linkage)
State the covalent interaction between lipoic acid and the enzyme dihydrolipoyl transacetylase (E2).