Regulation of the TCA Cycle Flashcards
Who first described the TCA cycle? Using what? What did he also contribute to?
- Krebs
- Using pigeon muscle
- Urea cycle
What is the underlying premise of the TCA cycle?
- Acetyl group of Acetyl-CoA is oxidized to CO2
- In the process, creates cellular respiration, energy released is conserved as NADH and FADH2
What is the most common input in the TCA cycle?
- Most common: Acetyl CoA
- Some amino acids can yield other intermediates
What mechanisms yield Acetyl-CoA?
- B-oxidation
- Amino acid catabolism (Leucine, threonine)
How many carbons does Acetyl-CoA contain? What about Pyruvate?
- Acetyl-CoA: 2
- Pyruvate: 3
Which enzyme catalyzes the conversion of Pyruvate to Acetyl-CoA? What type of reaction is it?
- Pyruvate Dehydrogenase Complex (PDC)
- IRREVERSIBLE oxidative decarboxylation
What is removed in the generation of Acetyl-CoA from Pyruvate?
Removal of hydrogen (oxidation) and removal of carbon (decarboxylation)
What is generated in the conversion of Pyruvate to Acetyl-CoA?
NADH
What is the composition of Coenzyme A?
- Adenine + Ribose-phosphate: Phosphoadenosine diphosphate
- Vitamin B5
- Reactive thiol group (CoA-SH)
What is the first enzyme of PDC?
E1: Pyruvate dehydrogenase
What is the second enzyme of PDC?
E2: Dihydrolipoyl transacetylase
What is the third enzyme of PDC?
E3: Dihydrolipoyl dehydrogenase
What are the co-substrates of PDC?
- CoA-SH
- NAD
What are the prosthetic groups of PDC?
- Thiamine pyrophosphate (TPP)
- Lipoate
- FAD
Which B vitamin composes TPP?
Thiamine (B1)
Which B vitamin composes FAD?
Riboflavin (B2)
Which B vitamin composes NAD?
Niacin (B3)
Which B vitamin composes CoA?
Pantothenic acid (B5)
What happens in pyruvate dehydrogenase (E1)
1) Pyruvate attaches to TPP on E1, forming a hydroxyethal TPP
2) CO2 is removed
What happens in dihydrolipoyl transacetylase (E2)
1) Lipoate is attached to Lysine on E2, which forms a lipoyllysine
2) Lipoyllysine is acetylated (from its oxidized form) when the acetyl group of Pyruvate is transferred
3) Lipoyllysine is changed to its reduced form when the acetyl group is transferred to CoA, forming Acetyl-CoA
What happens in dihydrolipoyl dehydrogenase (E3)?
1) FADH2 produced is oxidized to FAD
2) NAD is reduced to NADH
Which compounds in PDC are permanently bound? What are they called?
- Prosthetic groups
- TPP, lipoate, FAD
How many intermediates are there in the TCA cycle? How many enzymatic reactions?
- 9 intermediates
- 8 enzymatic reactions
How many carbons does oxaloacetate contain? What does it combine with to form citrate?
- Oxaloacetate: 4C
- Acetyl-CoA: 2C
- Oxaloacetate + Acetyl-CoA = Citrate
How many TCA cycle passes are needed to form CO2 from 2C of Acetyl-CoA? At which steps?
- Steps 3 and 4
- 2 TCA cycle passes
Which enzyme catalyzes the condensation step of the TCA cycle? What does it form? From what?
- Enzyme: Citrate synthase
- Oxaloacetate + Acetyl-CoA = CItrate
What does citrate inhibit?
- PFK1 (glycolysis)
- ACC (FA synthesis)
Which components of the TCA cycle have 3 carboxyl groups?
- Isocitrate
- Citrate
Which enzyme catalyzes the hydration/dehydration step of the TCA cycle? What does it form? From what?
- Enzyme: Aconitase
- Citrate -> Isocitrate
Which enzyme catalyzes the oxidative decarboxylation step of the TCA cycle? What does it form? From what?
- Enzyme: Isocitrate dehydrogenase
- Isocitrate -> a-ketoglutarate
What does isocitrate dehydrogenase synthesize as a by-product?
NADH or NAPDH depending on the enzyme isoform
Which steps generate energy molecules in the TCA cycle?
- Isocitrate dehydrogenase (NADH)
- a-Ketoglutarate dehydrogenase complex (NADH)
- Succinyl-CoA Synthetase (GTP or ATP)
- Succinate dehydrogenase (FADH2)
- Malate dehydrogenase (NADH)
Which steps generate CO2 in the TCA cycle?
- Isocitrate dehydrogenase
- a-Ketoglutarate dehydrogenase complex
Which enzyme catalyzes the second oxidative decarboxylation step of the TCA cycle? What does it form? From what?
- Enzyme: a-ketoglutarate dehydrogenase complex
- a-Ketoglutarate –> Succinyl-CoA
Which enzyme of the TCA cycle is similar to the PDC?
a-Ketoglutarate dehydrogenase complex
Which enzyme catalyzes the substrate-level phosphorylation step of the TCA cycle? What does it form? From what?
- Enzyme: Succinyl-CoA Synthetase
- Succinyl-CoA –> Succinate
Differentiate the two isoforms of Succinyl-CoA synthetase.
o Isoform that has affinity for GDP synthesizes GTP
o Isoform that has affinity for ADP synthesizes ATP
Which enzyme can convert a GTP into an ATP (or vice-versa)?
Nucleoside diphosphate kinase
Differentiate synthetase and synthase.
o Synthetase: when synthesis involves an energy molecule
o Synthase: when synthesis does not involve energy
Differentiate kinase and phosphorylase.
o Kinase: when synthesis involves an energy molecule
o Phosphorylase: when synthesis does not involve energy
Which enzyme catalyzes the dehydrogenation step of the TCA cycle? What does it form? From what?
- Enzyme: Succinate dehydrogenase
- Succinate –> Fumarate
Which enzyme of the TCA cycle is the only one that is located on the inner wall of the mitochondria?
Succinate dehydrogenase
Which enzyme catalyzes the second hydration step of the TCA cycle? What does it form? From what?
- Enzyme: Fumarase
- Fumarate –> Malate (OH- is added, then H+ is added)
Which enzyme catalyzes the second dehydrogenation step of the TCA cycle? What does it form? From what?
- Enzyme: Malate dehydrogenase
- Malate –> Oxaloacetate
What are the products of the TCA cycle?
Acetyl-CoA = 2CO2 + 3 NADH + 1 FADH2 + 1 ATP/GTP
What is an amphibolic pathway? Give an example.
- Biochemical pathway that involves catabolism and anabolism
(hub of mechanisms) - TCA cycle
What is the PDC inhibited by?
Inhibited by ATP, Acetyl-CoA, NADH, fatty acids
What is the PDC activated by?
Activated by AMP, CoA, NAD+, Ca2+
What is citrate synthase inhibited by?
Inhibited by NADH, Succinyl-CoA, citrate, ATP
What is citrate synthase activated by?
Activated by ADP
What is isocitrate dehydrogenase inhibited by?
Inhibited by ATP
What is isocitrate dehydrogenase activated by?
Activated by Ca2+, ADP
What is the a-ketoglutarate dehydrogenase complex inhibited by?
Inhibited by Succinyl-CoA, NADH
What is the a-ketoglutarate dehydrogenase complex activated by?
Activated by Ca2+
Which steps are irreversible in the TCA cycle?
1, 3, 4