ATP SYNTHESIS

ATP SYNTHESIS
General ATP Beta and gamma more energy Oxidative phosphorylation more imp. role in ATP formation than substrate Mitochondria Structure Outer Membrane Permeable Inner membrane Cristae Impermeable Rich in proteins/enzymesETC and ATP synthase Electron Transport System Four Protein Complexes Complex I Enzyme NADH DEHYDROGENASE Energy DonorNADH Energy AcceptorCoQ (Complex III) Prosthetic GroupsFMN, Fe-S Complex II Enzyme SUCCINATE DEHYDROGENASE Energy DonorFADH2 Energy Acceptor CoQ (Complex III) Prosthetic GroupsFAD, Fe-S Complex III Complex IV Donates Electron to Oxygen-H2O Formation of Electrochemical Potential ETC complexes are electron transporters as well as H+ pumps Protons pumped from MatrixIntermembrane space Complex I-III=4 Complex IIIII=0 Complex IIIIV=2 Complex IVO2=4 NADH VIA COMPLEX 1 =10 Protons=2.5 ATP FADH2 VIA COMPLEX 2=6 protons=1.5 ATP

Establishes ELECTROCHEMCIAL POTENTIAL Matrix negative compared to intermembrane space Provides driving force of ATP synthesis ATP Synthase Fopore which protons return to matrix Proton current causes roation of Fo and gamma subunit of F1 Conformational change in F1 leads to three steps Binding of Pi and ADPalpha and beta pairs bind Synthesis of ATPeach ATP requires 4 protons Releasing Product Oxidative Phosphorylation Oxidation Coenzymes NADH or FADH2 oxidized via ETC-O2H2o Phosphorylation ADP + PiATP Electrochemical potential is driving force Transporters in the Mitochondrial Inner Membrane Adenosine Nucleotide Translocase (ANTIPORT) ATP into Matrix ADP out Phosphate Translocase (SYMPORT) Phosphate into Matrix H+ into matrix Summary of Oxidative Phosphorylation

ATP Synthesisinterconversion of energyoxidation reductionelectrochemical gradient drives ATP synthesis

Reducing Equivalents Defined as 1 Hydrogen Atom (1 proton+ 1 electron) Carriers NADNADH NADPNADPH FADFADH2 FMN

ETS, ATP SYNTHASE AND OXIDATIVE PHOSPHORYLATION

Reducing Equivalents generated by FuelsProtein, Fats, CHO Oxidation of food stored generates reducing equivalents and ATP synthesis Conversion to Acetyl CoA Oxidation of Acetyl CoA via TCA Generation of Reducing equivalents

Synthesis of ATP

Acetyl CoA is common catabolism product Fatty Acid-palmitate3 Sugar2 Ketone Body-acetoacetate2 AA-alanine1 Ethanol1

Reducing Equivalent Shuttle Systems

Carriers of Electrons and protons but not the NADH molecule to the matrix Malate/Aspartate Shuttle (LIVER) 1. NADH reduces oxaloacetate to malate Enzyme MALATE DEHYDROGENASE 2. Malate enters mitochondria MALATE/alpha-kg transporter 3. Malate re-oxidized to OAA in Mito matrix Produces NADH 4. NADH enters ETC2.5 ATPs 5. OAA transaminated to Aspartate Enzyme ASPARTATE-GLUTAMATE TRANSPORTER 6. Aspartate Transported back to cytosol ASPARTATE GLUTAMATE TRANSPORTER Glycerol-3 Phosphate Shuttle (BRAIN) Dihydroxyacetone Phosphate (DHAP) reduced to glycerol-3-phosphate By NADH produced in glycolysis G3P oxidized to DHAP in the inner mitochondrial membrane Enzyme G3P DEHYDROGENASE Transfers electrons to FAD to form FADH2 FADH2 passes electrons to Complex III1.5 ATP when oxidized

Regulation of Oxidative Phosphorylation

Role of ADP in Respiratory Control Speed of oxygen reduction (decline in O2 concentration) represents speed of oxidative phosphorylation SubstratesPyruvate and malate enhance ADPenhances rapidly ADP/ATP and NAD/NADH ratios control oxidative phosphorylation The rations of ADP/ATP and NAD/NADH are important regulatory factors of electron transport chain reaction, oxidative phosphorylation, and TCA cycle Uncoupling Phosphorylation from Electron Transport Chain

Disconnecting of ETC building of electrochemical potential and ATP synthesis=Uncoupler Examples DNP (dinitrophenol, a synthesized uncoupler) Phenolic Hydroxyl group that can be protonated/deprot in response to pH Intermebrane space high Hydrogen Ion concentrationprotonated Protonated=lipophilic-rapidly diffuses into matrix Matrixdeprotonated (-) Moves to the innermembrane space Counteracts the H+ pumps and disrupts electrochemical gradient Presnece of uncoupler ETC will proceed Electrochemical potential will be disrupted NO ATP synthesis Uncoupling Proteins (UCP) UCP1newborns inner mito membrane brown adipose Mediates protons flowing from the intermembrane space to the matrix Electrochemical potential used for heat generation Cold+ UCP1 via norepi, B-adrenergic ColdNEpiB-adrencAMPPKALipolysisFFAUCP-1

Tri-iodothyrine T3 affects expression of proteins involved in basal metabolic rate and thermo genesis UCP-1 and UCP-3 Sarco/Endoplasmic Recticulum Ca-ATPase (SERCA1 and 2) Na/K ATPase Inhibitors of ATP Synthase Oligomycin Binds enzyme prevents H+ through proton channel Does not inhibit respiration directly but prevents dual stimulation by ADP An uncoupler can still stimulate oxygen consumption in the presence of oligomyosin Conclusions from the Experiments (Oligomycin vs Uncoupler) The energy that regulates the coupled system The uncoupler eliminates regulation by energy needs, and the ETS consumes O2 trying to restore the proton gradient The proton gradient also regulates the coupled system

Oligomycin shuts off ATP synthase, the H+ concentration gradient increases, slowing the rate of ETS, until uncoupler dissipates gradient and the ETC continues unchecked The Coupling of Processes to body needs links metabolic pathways Interlocking regulation of glycolysis, pyruvate oxidation, the citric acid cycle, and oxidative phosphorylation by relative concentrations of ATP, ADP, AMP and by NADH High ATP (Low ADP, AMP) produces low rates of Glycolysis Pyruvate oxidation Acetate oxidate via TCA And oxidative phosphorylation All four are stimulated by increases in rate of ATP utilization and ADP, AMP, Pi levels Interlocking of glycolysis and TCA by citrateinhibits glycolysis Supplements the action of adenine nucleotide system Increased levels of NADH and Acetyl coA also inhibit the oxidation of pyruvate to acetyl CoA and high NADH/NAD+ ratios inhibit the dehydrogenases of the TCA