Flashcards on Glycolysis
Q: What does the term "glycolysis" mean and what occurs during this process?
A:
Derived from Greek: "Glykys" = sweet, "Lysis" = splitting
One glucose (6C) molecule is degraded into two pyruvate (3C) molecules
Q: What is the most common glycolytic pathway and who discovered it?
A:
Embden-Meyerhof-Parnas (EMP) pathway
Discovered by Gustav Embden, Otto Meyerhof, Jakub Karol Parnas
Q: Name other types of glycolytic pathways.
A:
Enter-Doudoroff pathway
Heterofermentative and homofermentative pathways
Q: Who discovered cell-free fermentation?
A:
Eduard Buchner
Q: Who were the main scientists involved in elucidating glycolysis?
A:
Otto Meyerhof
Gustav Embden
Jakub Karol Parnas
Louis Pasteur
Q: What is Pasteur’s effect?
A:
Aerobic growth requires less glucose than anaerobic conditions
Q: What did Buchner demonstrate in 1897?
A:
Glucose can be converted to ethanol using non-living yeast extract (enzymes)
Q: What were Harden and Young’s key discoveries (1905-1911)?
A:
Inorganic phosphate is required for fermentation
Yeast extract contains small coenzymes (Co-zymase) and larger enzymes (zymase)
ATP regulates glucose consumption during alcohol fermentation
Identified fructose 1,6-bisphosphate as a glycolytic intermediate
Q: What did inhibitor studies reveal about glycolysis intermediates?
A:
Iodoacetate causes accumulation of fructose 1,6-bisphosphate
Fluoride causes accumulation of 2-phosphoglycerate and 3-phosphoglycerate
Q: Who proposed the detailed step-by-step glycolysis pathway in the 1930s?
A:
Gustav Embden
Q: What is the overall free energy change (ΔG°) of glycolysis?
A:
ΔG° = -85 kJ/mol (exergonic, irreversible under standard conditions)
Q: What is the fate of glucose in living systems (energy-wise)?
A:
Complete oxidation: Glucose + 6O2 → 6CO2 + 6H2O, ΔG° = -2840 kJ/mol
Glycolysis: Glucose + 2NAD+ → 2Pyruvate + 2NADH + 2H+, ΔG° = -146 kJ/mol
Glycolysis releases 5.2% of total free energy of glucose
Q: Where does glycolysis occur in the cell?
A:
Cytosol
Q: What are the net products of glycolysis?
A:
2 ATP (net gain)
2 NADH + H+
2 pyruvate
Q: What are the two phases of glycolysis?
A:
Preparatory (investment) phase: ATP consumed
Payoff phase: ATP produced
Q: What is the energy balance sheet for glycolysis?
A:
Gains: 4 ATP, 2 NADH + H+, 2 pyruvate
Losses: 2 ATP, glucose, phosphate, NAD+ (recycled)
Net gain: +2 ATP
Q: Why are glycolytic intermediates phosphorylated?
A:
Phosphates give intermediates a net negative charge (trapped in cell)
Phosphates are essential for enzymatic conservation of energy (form high-energy
compounds)
Phosphate binding lowers activation energy and increases enzyme specificity
Most glycolytic enzymes require Mg2+ for activity
Q: What is the first step of glycolysis and its enzyme?
A:
Hexokinase catalyzes: Glucose + ATP → Glucose-6-phosphate + ADP
ATP binds as Mg2+ complex
Q: What is the difference between hexokinase and glucokinase?
A:
Not detailed in notes, but both phosphorylate glucose, with glucokinase (hexokinase IV)
having different regulatory properties
Q: What happens in the isomerization of glucose-6-phosphate?
A:
Enzyme: Phosphohexoisomerase/phosphoglucoisomerase
Glucose-6-phosphate ↔ Fructose-6-phosphate
Involves ring opening/closing, acid/base catalysis, and a cis-enediol intermediate
Q: Which enzyme catalyzes the rate-limiting step of glycolysis?
A:
Phosphofructokinase
Reaction: Fructose-6-phosphate + ATP → Fructose-1,6-bisphosphate + ADP
Q: What does aldolase do in glycolysis?
A:
Catalyzes: Fructose-1,6-bisphosphate → Dihydroxyacetone phosphate (DHAP) +
Glyceraldehyde-3-phosphate
Q: What is the role of triose phosphate isomerase?
A:
Catalyzes: DHAP ↔ Glyceraldehyde-3-phosphate
Proceeds via enediol intermediate, acid/base catalysis by Glu and His residues
2-phosphoglycolate is a transition state analog inhibitor
Q: Which glycolytic step produces NADH?
A:
Glyceraldehyde-3-phosphate dehydrogenase catalyzes:
Glyceraldehyde-3-phosphate + NAD+ + Pi → 1,3-bisphosphoglycerate + NADH + H+
Only step reducing NAD+ to NADH
Q: What happens during the phosphoglycerate kinase step?
A:
Catalyzes: 1,3-bisphosphoglycerate + ADP ↔ 3-phosphoglycerate + ATP
Reversible phosphate transfer, substrate-induced conformational change
Q: What is the function of phosphoglycerate mutase?
A:
Catalyzes: 3-phosphoglycerate ↔ 2-phosphoglycerate
Phosphate shifted from C3 to C2 via active-site His residues
Q: What reaction does enolase catalyze?
A:
2-phosphoglycerate → Phosphoenolpyruvate + H2O
Mg2+-dependent dehydration reaction, stabilizes enolate anion intermediate
Q: What is the final step of glycolysis and its enzyme?
A:
Pyruvate kinase catalyzes: Phosphoenolpyruvate + ADP → Pyruvate + ATP
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Flashcards on Pyruvate Dehydrogenase Complex, Krebs Cycle, and Related
Concepts
Q: What is Mitochondrial Pyruvate Carrier 2 (MPC2) and its function?
A:
Also known as brain protein 44 (BRP44)
Encoded by the MPC2 gene in humans
Part of the mitochondrial pyruvate carrier (MPC) protein family
Involved in transport of pyruvate across the inner mitochondrial membrane [1]
Q: What are the three enzymes of the pyruvate dehydrogenase complex and their prosthetic
groups/cofactors?
A:
E1: Pyruvate dehydrogenase (uses thiamine pyrophosphate, TPP)
E2: Dihydrolipoyl transacetylase (uses lipoamide and coenzyme A/CoASH)
E3: Dihydrolipoyl dehydrogenase (uses flavin adenine dinucleotide, FAD, and nicotinamide
adenine dinucleotide, NAD+) [1]
Q: What are the five steps of the pyruvate dehydrogenase reaction?
A:
Step A: Pyruvate is decarboxylated by E1 with TPP
Step B: Acetyl group transferred to lipoamide, forming hydroxyethyl-TPP, releasing CO2
Step C: E2 oxidizes hydroxyethyl- to acetyl-, transfers acetyl- to CoA, forming acetyl-CoA
Step D: E3 oxidizes dihydrolipoamide back to lipoamide
Step E: NAD+ is reduced to NADH as a side reaction [1]
Q: What is pyruvate dehydrogenase deficiency (PDCD) and its main features?
A:
One of the most common neurodegenerative disorders linked to abnormal mitochondrial
metabolism
X-linked disease with heterogeneous clinical and biochemical presentations
Two forms: metabolic (lactic acidosis) and neurological (hypotonia, poor feeding, lethargy,
brain abnormalities, seizures, spasms)
Can progress to mental retardation, microcephaly, blindness, spasticity
Females with residual activity: few symptoms
Females with little/no activity: major brain abnormalities, atrophy
Males with little/no activity: die in utero due to insufficient ATP in brain cells [1]
Q: Who proposed the citric acid (Krebs) cycle and what was the contribution of coenzyme A
discovery?
A:
Proposed by Sir Hans Adolf Krebs in 1937 (Nobel Prize 1953)
Discovery of coenzyme A in 1945 by Fritz Lipmann and Nathan Kaplan completed
understanding of the cycle [1]
Q: What is the first step of the Krebs cycle and the enzyme involved?
A:
Condensation of acetyl-CoA with oxaloacetate to form citrate
Catalyzed by citrate synthase
Oxaloacetate binds first, causing conformational change for acetyl-CoA binding [1]
Q: How is citrate converted to isocitrate?
A:
Citrate is isomerized to isocitrate via cis-aconitate intermediate
Catalyzed by aconitase (contains iron-sulfur center)
Reaction is reversible, pulled forward in cells by rapid consumption of isocitrate [1]
Q: Describe the oxidation of isocitrate in the Krebs cycle.
A:
Isocitrate dehydrogenase catalyzes oxidative decarboxylation to α-ketoglutarate
Requires Mn2+ in active site
Two forms: NAD+-dependent (mitochondrial matrix, TCA cycle) and NADP+-dependent
(matrix and cytosol, NADPH generation) [1]
Q: What happens during the conversion of α-ketoglutarate to succinyl-CoA?
A:
Catalyzed by α-ketoglutarate dehydrogenase complex
Another oxidative decarboxylation
NAD+ as electron acceptor, CoA as succinyl group carrier
Energy conserved in thioester bond of succinyl-CoA [1]
Q: How is succinyl-CoA converted to succinate?
A:
Catalyzed by succinyl-CoA synthetase (succinic thiokinase)
Hydrolysis of thioester bond drives synthesis of GTP or ATP
Net ΔG'° is -2.9 kJ/mol [1]
Q: What is the role of succinate dehydrogenase in the Krebs cycle?
A:
Oxidizes succinate to fumarate
Flavoprotein tightly bound to mitochondrial inner membrane (eukaryotes)
Contains iron-sulfur clusters and covalently bound FAD [1]
Q: How is fumarate converted to malate?
A:
Hydration of fumarate to L-malate
Catalyzed by fumarase (fumarate hydratase)
Highly stereospecific for trans double bond of fumarate [1]
Q: What is the final step of the Krebs cycle?
A:
Oxidation of L-malate to oxaloacetate
Catalyzed by NAD-linked L-malate dehydrogenase
Reaction equilibrium favors malate, but pulled forward by citrate synthase reaction [1]
Q: How does the citric acid cycle contribute to ATP production?
A:
Directly generates 1 ATP (or GTP) per turn (succinyl-CoA to succinate)
Four oxidation steps produce NADH and FADH2, which drive ATP synthesis via oxidative
phosphorylation [1]
Q: What is the ATP yield from complete aerobic oxidation of glucose?
A:
As many as 32 ATP per glucose
2.5 ATP per NADH, 1.5 ATP per FADH2
Includes both pyruvate oxidation and citric acid cycle, with electrons transferred to O2 via
oxidative phosphorylation [1]
Q: What is the malate-aspartate shuttle and where is it used?
A:
Transports reducing equivalents from cytosolic NADH into mitochondrial matrix
Used in liver, kidney, and heart
Steps: NADH reduces oxaloacetate to malate (cytosol) → malate enters matrix → malate
oxidized to oxaloacetate (matrix) → oxaloacetate transaminated to aspartate → aspartate
exits to cytosol → oxaloacetate regenerated [1]
Q: What is the glycerol 3-phosphate shuttle and where does it operate?
A:
Moves reducing equivalents from cytosol to mitochondrial matrix
Operates in skeletal muscle and brain
Dihydroxyacetone phosphate reduced to glycerol 3-phosphate (cytosol), which is re-
oxidized on outer face of inner membrane, transferring electrons to ubiquinone [1]
Q: What are anaplerotic reactions and why are they important?
A:
Reactions that replenish TCA cycle intermediates depleted for biosynthesis
Maintain constant concentrations of TCA intermediates
Allow dynamic balance between biosynthetic use and replenishment [1]
Q: How is the TCA cycle regulated?
A:
Regulation at two main entry points: conversion of pyruvate to acetyl-CoA (pyruvate
dehydrogenase complex) and entry of acetyl-CoA into cycle (citrate synthase)
Also regulated at isocitrate dehydrogenase and α-ketoglutarate dehydrogenase steps [1]
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1. [Link]
