⚗️Biological Chemistry II Unit 6 – Electron Transport & Oxidative Phosphorylation

Key Concepts and Terminology

• Electron transport chain (ETC) consists of a series of protein complexes embedded in the inner mitochondrial membrane that facilitate the transfer of electrons from reduced electron carriers to oxygen
• Oxidative phosphorylation couples the energy released from electron transport to the synthesis of ATP through the formation of a proton gradient across the inner mitochondrial membrane
• Chemiosmotic theory explains the mechanism by which ATP is generated through the coupling of electron transport and proton gradient formation
• Proton motive force (PMF) represents the energy stored in the proton gradient, which drives ATP synthesis via ATP synthase
• Redox reactions involve the transfer of electrons from a reduced electron donor to an oxidized electron acceptor, releasing energy in the process
• Electron carriers, such as NADH and FADH2, donate electrons to the ETC, initiating the process of electron transport
• Mitochondrial respiratory complexes (I, II, III, and IV) are protein complexes that facilitate the transfer of electrons along the ETC
• ATP synthase (Complex V) is an enzyme that catalyzes the synthesis of ATP using the energy stored in the proton gradient

Electron Transport Chain Components

• Complex I (NADH dehydrogenase) accepts electrons from NADH and transfers them to ubiquinone (coenzyme Q10), pumping protons across the inner mitochondrial membrane
• Complex II (succinate dehydrogenase) is an enzyme that participates in both the citric acid cycle and the ETC, transferring electrons from succinate to ubiquinone without pumping protons
• Ubiquinone (coenzyme Q10) is a lipid-soluble electron carrier that shuttles electrons between Complexes I, II, and III
• Complex III (cytochrome bc1 complex) transfers electrons from ubiquinone to cytochrome c, pumping protons across the inner mitochondrial membrane
• Cytochrome c is a water-soluble electron carrier that shuttles electrons from Complex III to Complex IV
• Complex IV (cytochrome c oxidase) is the terminal enzyme of the ETC, transferring electrons from cytochrome c to oxygen, pumping protons across the inner mitochondrial membrane
• Supercomplexes are higher-order assemblies of individual respiratory complexes that enhance electron transport efficiency and minimize reactive oxygen species (ROS) production

Stages of Electron Transport

• Stage 1: Electron entry
  • NADH donates electrons to Complex I, while FADH2 donates electrons to Complex II
  • Electrons from both complexes are transferred to ubiquinone
• Stage 2: Electron shuttling
  • Reduced ubiquinone (ubiquinol) carries electrons to Complex III
  • Complex III transfers electrons from ubiquinol to cytochrome c, pumping protons across the inner mitochondrial membrane
• Stage 3: Electron transfer to oxygen
  • Cytochrome c shuttles electrons from Complex III to Complex IV
  • Complex IV transfers electrons from cytochrome c to oxygen, the terminal electron acceptor, forming water
  • Protons are pumped across the inner mitochondrial membrane during this process
• Proton pumping occurs at Complexes I, III, and IV, contributing to the formation of the proton gradient
• The flow of electrons through the ETC is accompanied by a decrease in redox potential, releasing energy that is used to pump protons across the inner mitochondrial membrane

Proton Gradient Formation

• Proton pumping by Complexes I, III, and IV results in the accumulation of protons in the intermembrane space of the mitochondria
• The inner mitochondrial membrane is impermeable to protons, maintaining the proton gradient
• The proton gradient generates an electrochemical potential difference across the inner mitochondrial membrane, known as the proton motive force (PMF)
• The PMF consists of two components:
  • Electrical component (ΔΨ): Difference in electrical charge across the membrane
  • Chemical component (ΔpH): Difference in proton concentration across the membrane
• The energy stored in the PMF is used to drive ATP synthesis via ATP synthase
• The magnitude of the PMF is approximately 200 mV, which is sufficient to drive ATP synthesis
• The proton gradient is maintained by the continuous pumping of protons by the ETC complexes and the controlled flow of protons back into the matrix through ATP synthase

ATP Synthesis via ATP Synthase

• ATP synthase (Complex V) is a molecular machine that catalyzes the synthesis of ATP using the energy stored in the proton gradient
• The enzyme consists of two main components:
  • F0 subunit: Embedded in the inner mitochondrial membrane, forms a proton channel
  • F1 subunit: Protrudes into the mitochondrial matrix, contains the catalytic sites for ATP synthesis
• Protons flow down their electrochemical gradient through the F0 subunit, causing it to rotate
• The rotation of the F0 subunit drives conformational changes in the F1 subunit, leading to ATP synthesis
• The binding change mechanism explains the process of ATP synthesis:
  • ADP and inorganic phosphate (Pi) bind to the catalytic sites on the F1 subunit
  • Proton flow through the F0 subunit causes rotation, leading to conformational changes in the F1 subunit
  • The conformational changes result in the formation of ATP from ADP and Pi
  • ATP is released from the catalytic sites, and the process repeats
• The number of protons required for the synthesis of one ATP molecule varies between species, typically ranging from 2.7 to 3.3 protons per ATP

Chemiosmotic Theory

• Chemiosmotic theory, proposed by Peter Mitchell, explains the coupling of electron transport and ATP synthesis through the formation of a proton gradient
• The theory states that the energy released from electron transport is used to pump protons across the inner mitochondrial membrane, creating an electrochemical gradient (PMF)
• The PMF drives the synthesis of ATP via ATP synthase, as protons flow down their electrochemical gradient
• The key components of the chemiosmotic theory are:
  • Electron transport chain: Transfers electrons from reduced electron carriers to oxygen, releasing energy
  • Proton pumping: ETC complexes use the released energy to pump protons across the inner mitochondrial membrane
  • Proton gradient: The accumulation of protons in the intermembrane space creates an electrochemical gradient (PMF)
  • ATP synthase: Utilizes the energy stored in the PMF to synthesize ATP as protons flow back into the matrix
• The chemiosmotic theory provides a unified explanation for the mechanism of ATP synthesis in mitochondria, chloroplasts, and bacteria
• Experimental evidence supporting the chemiosmotic theory includes:
  • Inhibition of electron transport or proton pumping leads to a decrease in ATP synthesis
  • Uncouplers (e.g., 2,4-dinitrophenol) that dissipate the proton gradient also inhibit ATP synthesis

Regulation and Inhibition

• Electron transport and oxidative phosphorylation are regulated to meet the energy demands of the cell
• Substrate availability (NADH and FADH2) is a key factor in regulating the rate of electron transport
  • Increased substrate availability (e.g., during high energy demand) enhances electron transport and ATP synthesis
  • Decreased substrate availability (e.g., during low energy demand) slows down electron transport and ATP synthesis
• Allosteric regulation of ETC complexes and ATP synthase fine-tunes the rate of electron transport and ATP synthesis
  • ATP acts as an allosteric inhibitor of Complex IV, reducing electron transport when ATP levels are high
  • ADP acts as an allosteric activator of ATP synthase, stimulating ATP synthesis when ADP levels are high
• Respiratory control is the regulation of electron transport by the availability of ADP and Pi
  • High ADP and Pi levels stimulate electron transport and ATP synthesis
  • Low ADP and Pi levels slow down electron transport and ATP synthesis
• Inhibitors of electron transport and oxidative phosphorylation can be used to study the mechanism and regulation of these processes
  • Rotenone inhibits Complex I by blocking electron transfer from NADH to ubiquinone
  • Antimycin A inhibits Complex III by blocking electron transfer from cytochrome b to cytochrome c1
  • Cyanide and carbon monoxide inhibit Complex IV by binding to the heme group of cytochrome a3
  • Oligomycin inhibits ATP synthase by blocking the proton channel in the F0 subunit

Real-World Applications and Disorders

• Understanding the mechanisms of electron transport and oxidative phosphorylation is crucial for developing treatments for mitochondrial disorders
• Mitochondrial disorders are a group of genetic diseases characterized by defects in electron transport, oxidative phosphorylation, or mitochondrial DNA
  • Examples include Leigh syndrome, MELAS syndrome, and Kearns-Sayre syndrome
  • Symptoms can vary depending on the specific mutation and affected tissues but often include muscle weakness, neurological problems, and developmental delays
• Aging and age-related diseases have been linked to mitochondrial dysfunction and oxidative stress
  • Mitochondrial DNA mutations accumulate over time, leading to decreased efficiency of electron transport and ATP synthesis
  • Increased production of reactive oxygen species (ROS) by dysfunctional mitochondria can damage cellular components and contribute to the aging process
• Targeting electron transport and oxidative phosphorylation is a potential strategy for treating cancer
  • Cancer cells often exhibit altered mitochondrial function and increased reliance on glycolysis for energy production (Warburg effect)
  • Drugs that inhibit ETC complexes or disrupt the proton gradient (e.g., metformin, 2-deoxyglucose) are being investigated as potential cancer therapies
• Mitochondrial function and oxidative phosphorylation are important factors in the study of exercise physiology and athletic performance
  • Endurance training increases mitochondrial content and efficiency, enhancing the capacity for ATP production
  • Nutritional strategies (e.g., ketogenic diets, creatine supplementation) can influence mitochondrial function and energy production in athletes