Passive Transport Requires Atp
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Cellular Biology 1
Cellular biology 1 INTRODUCTION • Specialized intracellular membrane-bound organelles (Fig. 1.2), such as mitochondria, Golgi apparatus, endoplasmic reticulum (ER). This chapter is an overview of eukaryotic cells, addressing • Large size (relative to prokaryotic cells). their intracellular organelles and structural components. A basic appreciation of cellular structure and function is important for an understanding of the following chapters’ information concerning metabolism and nutrition. For fur- ther detailed information in this subject area, please refer to EUKARYOTIC ORGANELLES a reference textbook. Nucleus The eukaryotic cell The nucleus is surrounded by a double membrane (nuclear Humans are multicellular eukaryotic organisms. All eukary- envelope). The envelope has multiple pores to allow tran- otic organisms are composed of eukaryotic cells. Eukaryotic sit of material between the nucleus and the cytoplasm. The cells (Fig. 1.1) are defined by the following features: nucleus contains the cell’s genetic material, DNA, organized • A membrane-limited nucleus (the key feature into linear structures known as chromosomes. As well as differentiating eukaryotic cells from prokaryotic cells) chromosomes, irregular zones of densely staining material that contains the cell’s genetic material. are also present. These are the nucleoli, which are responsible Inner nuclear Nucleus membrane Nucleolus Inner Outer Outer mitochondrial nuclear mitochondrial membrane membrane membrane Ribosome Intermembrane space Chromatin Mitochondrial Rough matrix Mitochondrial Nuclear endoplasmic ribosome pore reticulum Crista Mitochondrial mRNA Smooth Vesicle endoplasmic Mitochondrion Circular reticulum mitochondrial Proteins of the DNA Vesicle budding electron transport off rough ER Vesicles fusing system with trans face of Cytoplasm Golgi apparatus ‘Cis’ face + discharging protein/lipid Golgi apparatus ‘Trans’ face Lysosome Vesicles leaving Golgi with modified protein/lipid cargo Cell membrane Fig. -
Passive and Active Transport
Passive and Active Transport 1. Thermodynamics of transport 2. Passive-mediated transport 3. Active transport neuron, membrane potential, ion transport Membranes • Provide barrier function – Extracellular – Organelles • Barrier can be overcome by „transport proteins“ – To mediate transmembrane movements of ions, Na+, K+ – Nutrients, glucose, amino acids etc. – Water (aquaporins) 1) Thermodynamics of Transport • Aout <-> Ain (ressembles a chemical equilibration) o‘ • GA - G A = RT ln [A] • ∆GA = GA(in) - GA(out) = RT ln ([A]in/[A]out) • GA: chemical potential of A o‘ • G A: chemical potential of standard state of A • If membrane has a potential, i.e., plasma membrane: -100mV (inside negative) then GA is termed the electrochemical potential of A Two types of transport across a membrane: o Nonmediated transport occurs by passive diffusion, i.e., O2, CO2 driven by chemical potential gradient, i.e. cannot occur against a concentration gradient o Mediated transport occurs by dedicated transport proteins 1. Passive-mediated transport/facilitated diffusion: [high] -> [low] 2. Active transport: [low] -> [high] May require energy in form of ATP or in form of a membrane potential 2) Passive-mediated transport Substances that are too large or too polar to diffuse across the bilayer must be transported by proteins: carriers, permeases, channels and transporters A) Ionophores B) Porins C) Ion Channels D) Aquaporins E) Transport Proteins A) Ionophores Organic molecules of divers types, often of bacterial origin => Increase the permeability of a target membrane for ions, frequently antibiotic, result in collapse of target membrane potential by ion equilibration 1. Carrier Ionophore, make ion soluble in membrane, i.e. valinomycin, 104 K+/sec 2. -
Evidence for a Respiratory Chain in the Chloroplast
Proc. NatL Acad. Sci. USA Vol. 79, pp. 4352-4356, July 1982 Cell Biology Evidence for a respiratory chain in the chloroplast (photosynthesis/respiration/starch degradation/evolution) PIERRE BENNOUN Institut de Biologie Physico-Chimique, 13, rue Pierre et Marie Curie, 75005, Paris, France Communicated by Pierre Joliot, April 12, 1982 ABSTRACT Evidence is given for the existence ofan electron in 20 ml of 20 mM N-tris(hydroxymethyl)methylglycine(Tri- transport pathway to oxygen in the thylakoid membranes ofchlo- cine)/KOH, pH 7.8/10 mM NaCl/10 mM MgCl2/1 mM K2- roplasts (chlororespiration). Plastoquinone is shown to be a redox HPO4/0.1 M sucrose/5% Ficoll. The cell suspension was carrier common to both photosynthetic and chlororespiratory passed through a Yeda press operated at 90 kg/cm2, diluted pathways. It is shown that, in dark-adapted chloroplasts, an elec- with 200 ml of Ficoll-lacking buffer, and centrifuged, and the trochemical gradient is built up across the thylakoid membrane pellet was suspended in the same buffer. by transfer of electrons through the chlororespiratory chain as Chlorophyll fluorescence kinetics and luminescence mea- well as by reverse functioning of the chloroplast ATPases. It is surements were performed as described (9). proposed that these mechanisms ensure recycling ofthe ATP and NAD(P)H generated by the glycolytic pathway converting starch into triose phosphates. Chlororespiration is thus an 02-uptake RESULTS process distinct from photorespiration and the Mehler reaction. The plastoquinone (PQ) pool ofchloroplast is a redox carrier of The evolutionary significance of chlororespiration is discussed. the photosynthetic electron transport chain. -
The Electrochemical Gradient of Protons and Its Relationship to Active Transport in Escherichia Coli Membrane Vesicles
Proc. Natl. Acad. Sci. USA Vol. 73, No. 6, pp. 1892-1896, June 1976 Biochemistry The electrochemical gradient of protons and its relationship to active transport in Escherichia coli membrane vesicles (flow dialysis/membrane potential/energy transduction/lipophilic cations/weak acids) SOFIA RAMOS, SHIMON SCHULDINER*, AND H. RONALD KABACK The Roche Institute of Molecular Biology, Nutley, New Jersey 07110 Communicated by B. L. Horecker, March 17, 1976 ABSTRACT Membrane vesicles isolated from E. coli gen- presence of valinomycin), a respiration-dependent membrane erate a trans-membrane proton gradient of 2 pH units under potential (AI, interior negative) of approximately -75 mV in appropriate conditions when assayed by flow dialysis. Using E. coli membrane vesicles has been documented (6, 13, 14). the distribution of weak acids to measure the proton gradient (ApH) and the distribution of the lipophilic cation triphenyl- Moreover it has been shown that the potential causes the ap- methylphosphonium to measure the electrical potential across pearance of high affinity binding sites for dansyl- and azido- the membrane (AI), the vesicles are shown to generate an phenylgalactosides on the outer surface of the membrane (4, electrochemical proton gradient (AiH+) of approximately -180 15) and that the potential is partially dissipated as a result of mV at pH 5.5 in the presence of ascorbate and phenazine lactose accumulation (6). Although these findings provide ev- methosulfate, the major component of which is a ApH of about idence for the chemiosmotic hypothesis, it has also been dem- -110 mV. As external pH is increased, ApH decreases, reaching o at pH 7.5 and above, while AI remains at about -75 mV and onstrated (6, 16) that vesicles are able to accumulate lactose and internal pH remains at pH 7.5. -
Photosynthesis and Respiration
18 Photosynthesis and Respiration ATP is the energy currency of the cell Goal To understand how energy from sunlight is harnessed to Cells need to carry out many reactions that are energetically unfavorable. generate chemical energy by photosynthesis and You have seen some examples of these non-spontaneous reactions in respiration. earlier chapters: the synthesis of nucleic acids and proteins from their corresponding nucleotide and amino acid building blocks and the transport Objectives of certain ions against concentration gradients across a membrane. In many cases, unfavorable reactions like these are coupled to the hydrolysis of ATP After this chapter, you should be able to: in order to make them energetically favorable under cellular conditions; we • Explain the concepts of oxidation and have learned that for these reactions the free energy released in breaking reduction. the phosphodiester bonds in ATP exceeds the energy consumed by the • Explain how light energy generates an uphill reaction such that the sum of the free energy of the two reactions is electrochemical gradient. negative (ΔG < 0). To perform these reactions, cells must then have a way • Explain how an electrochemical of generating ATP efficiently so that a sufficient supply is always available. gradient generates chemical energy. The amount of ATP used by a mammalian cell has been estimated to be on the order of 109 molecules per second. In other words, ATP is the principal • Explain how chemical energy is harnessed to fix carbon dioxide. energy currency of the cell. • Explain how glucose is used to generate How does the cell produce enough ATP to sustain life and what is the source ATP anaerobically. -
3. Transport Can Be Active Or Passive. •Passive Transport Is Movement
3. Transport can be active or passive. F 6-3 Taiz. Microelectrodes are used to measure membrane •Passive transport is movement down an electrochemical potentials across cell membrane gradient. •Active transport is movement against an electrochemical gradient. What is an electrochemical gradient? How is it formed? Passive and active transport of ions result in electric potential difference across membranes. •Movement of an uncharged mol Is dependent on conc. gradient alone. •Movement of an ion depends on the electric gradient and the conc. gradient. •Diffusion potential- Pump potential- How do you know if an ion is moving uphill or downhill? Nernst Eq What is the driving force for uphill movement? A) ATP ; b) H+ gradient 6-5. Pump potential and diffusion potential. How can we determine whether an ion moves in or out by active or passive transport? Nernst equation states that at equilibrium the difference in concentration of an ion between two compartments is balanced by the voltage difference. Thus it can predict the ion conc at equilibrium at a certain ΔE. Very useful to predict active or passive transport of an ion. Fig. 6-4, Taiz. Passive and active transporters. Tab 6-1, Taiz . Using the Nernst equation to predict ion conc. at equilibrium when the Cell electrical potential, Δψ = -110 mV ---------------------------------------------------------------------------------------- Ext Conc. Ion Internal concentration (mM) Summary: In general observed Nernst (Predicted) ---------------------------------------------------------------------------------------- Cation uptake: passive 1 mM K+ 75 mM 74 Cation efflux: active 1 mM Na+ 8 mM 74 1 mM Ca2+ 2 mM 5,000 Anion uptake: active 0.2 mM Mg2+ 3 1,340 Anion release: passive - 2 mM NO3 5 mM 0.02 1 Cl- 10 mM 0.01 - 1H2PO4 21 0.01 ---------------------------------------------------------------------------------------- 1 6-10. -
Molecular Biology of the Cell 6Th Edition
753 CHAPTER Energy Conversion: Mitochondria and Chloroplasts 14 To maintain their high degree of organization in a universe that is constantly drift- IN THIS CHAPTER ing toward chaos, cells have a constant need for a plentiful supply of ATP, as we have explained in Chapter 2. In eukaryotic cells, most of the ATP that powers life THE MITOCHONDRION processes is produced by specialized, membrane-enclosed, energy-converting organelles. Tese are of two types. Mitochondria, which occur in virtually all cells THE PROTON PUMPS OF THE of animals, plants, and fungi, burn food molecules to produce ATP by oxidative ELECTRON-TRANSPORT CHAIN phosphorylation. Chloroplasts, which occur only in plants and green algae, har- ness solar energy to produce ATP by photosynthesis. In electron micrographs, the ATP PRODUCTION IN most striking features of both mitochondria and chloroplasts are their extensive MITOCHONDRIA internal membrane systems. Tese internal membranes contain sets of mem- brane protein complexes that work together to produce most of the cell’s ATP. In CHLOROPLASTS AND bacteria, simpler versions of essentially the same protein complexes produce ATP, PHOTOSYNTHESIS but they are located in the cell’s plasma membrane (Figure 14–1). Comparisons of DNA sequences indicate that the energy-converting organ- THE GENETIC SYSTEMS elles in present-day eukaryotes originated from prokaryotic cells that were endo- OF MITOCHONDRIA AND cytosed during the evolution of eukaryotes (discussed in Chapter 1). This explains CHLOROPLASTS why mitochondria and chloroplasts contain their own DNA, which still encodes a subset of their proteins. Over time, these organelles have lost most of their own genomes and become heavily dependent on proteins that are encoded by genes in the nucleus, synthesized in the cytosol, and then imported into the organelle. -
Electron Transport Generates a Proton Gradient Across the Membrane
Electron Transport Generates a Proton Gradient Across the Membrane Each of respiratory enzyme complexes couples the energy released by electron transfer across it to an uptake of protons from water in the mitochondrial matrix, accompanied by the release of protons on the other side of the membrane into the intramembrane space. As result, the energetically favorable flow of electrons along the electron- transport chain pumps protons across the membrane out of the matrix. This event creates electrochemical protons across the inner membrane. The Proton Gradient Drives ATP Synthesis The electrochemical proton gradient across the inner mitochondrial membrane is used to drive ATP synthesis in the process of oxidative Phosphorylation. The device that makes this possible is a large membrane-bound enzyme called ATP synthase. This enzymes creates a hydrophilic pathway across the inner mitochondrial membrane that allows protons to follow down their electrochemical gradient. As these ions thread their way through the ATP synthase, they are used to drive the energetically unfavorable reaction between ADP and Pi. 1 Proton Gradients Produce Most of the Cell’s ATP Glycolysis alone produces a net yield of two molecules of ATP for every molecule of glucose, which is the total energy yield for the fermentation process that occur in the absence of oxygen. In contrast, during the oxidative Phosphorylation each pair of electrons donated by NADH produced mitochondria is thought to provide energy for the formation of the about 2.5 molecules of ATP, once one includes the energy needed for transporting this ATP to cytosol. Oxidative Phosphorylation also produces 1.5 ATP molecules per electron pair of FADH2, or from the NADH molecules produced by glycolysis in the cytosol. -
A) Thylakoid Membrane
1)Which incorrectly matches process and location? a) Oxygen gas is produced—the thylakoid space b) Activated chlorophyll donates an electron—the thylakoid membranes c) NADPH is oxidized to NADP+—the stroma d) ATP is produced—the intermembrane space e) Rubisco catalyzes carbon fixation—the stroma 1)Which incorrectly matches process and location? a) Oxygen gas is produced—the thylakoid space b) Activated chlorophyll donates an electron—the thylakoid membranes c) NADPH is oxidized to NADP+—the stroma d) ATP is produced—the intermembrane space e) Rubisco catalyzes carbon fixation—the stroma 2)Of these events from the light reactions, which occurs first? a) Light-induced reduction of the primary electron acceptor in the reaction center of PS II. b) While being split, electrons are taken out of water. c) Donation of electrons from reduced Pq to the cytochrome complex. d) Acceptance of electrons by Pc from the cytochrome complex. e) Pq gets electrons from the reduced primary electron acceptor of PS II. 2)Of these events from the light reactions, which occurs first? a) Light-induced reduction of the primary electron acceptor in the reaction center of PS II. b) While being split, electrons are taken out of water. c) Donation of electrons from reduced Pq to the cytochrome complex. d) Acceptance of electrons by Pc from the cytochrome complex. e) Pq gets electrons from the reduced primary electron acceptor of PS II. 3)When donating its activated electron, the chlorophyll in photosystem II is a very powerful oxidizing agent. This is best shown by its ability to a) make use of a proton electrochemical gradient to drive the formation of ATP. -
Generation of an Electrochemical Proton Gradient in Streptococcus Cremoris by Lactate Efflux (Fermentation/Transport) ROEL OTTO, ANTON S
Proc. Natl. Acad. Sci. USA Vol. 77, No. 9, pp. 5502-5506, September 1980 Microbiology Generation of an electrochemical proton gradient in Streptococcus cremoris by lactate efflux (fermentation/transport) ROEL OTTO, ANTON S. M. SONNENBERG, HANS VELDKAMP, AND WIL N. KONINGS* Department of Microbiology, Biological Centre, University of Groningen, Kerklaan 30, 9751 NN Haren, The Netherlands Communicated by Peter Mitchell, June 9, 1980 ABSTRACT Recently an energy-recycling model was pro- solute will occur when [(n - 1)AT - nZApH] < -Z posed that postulates the generation of an electrochemical log(A-/A-,) and the energy of the solute gradient then will be gradient in fermentative bacteria by carrier-mediated excretion into of the of metabolic end products in symport with protons. In this paper converted energy electrochemical proton gra- experimental support for this model is given. In batch cultures dient. of Streptococcus cremoris with glucose as the sole energy source During fermentation, excretion of metabolic end products the maximal specific growth rate decreased by 30% when the via a carrier in symport with proton(s) can occur only when the external lactate concentration was decreased from 50 to 90 mM. outwardly directed driving force supplied by the chemical end In the same range of external lactate concentrations the molar product gradient exceeds the inwardly directed driving force growth yield Y for glucose as measured in energy-limited che- supplied by the electrochemical gradient. The excretion of end mostat cultures also showed a 30% drop. From ynaxtose values of S. cremoris grown in the presence and absence of added lactate products will then lead to the generation of an electrochemical it was calculated that the net energy ain from the lactate efflux gradient. -
Principle of Chemiosmotic Mechanism Tianyi Shi 2019-10-15
Principle of chemiosmotic mechanism Tianyi Shi 2019-10-15 1 SAQ Explain the difference between electrochemical gradient, pH gradient and membrane poten- tials. How can they be measured and how are these related to chemiosmotic mechanism. The electrochemical gradient describes the difference in chemical stability of ions (typically protons), between two aqueous compartments separated by a non-conducting and non-permeable membrane (typically a energy- transducing membrane such as mitochondrial inner membrane). It is quantified by the difference in Gibbs’ free energy, ∆G, or its molar equivalent, chemical potential, ∆µ, of the ion species (proton) between the two compartments. Usually, electrochemical gradient is used in studying processes coupled with H+ movement across a membrane, where one side is positively charged and has a high H+ and the other side with -ve charge and low H+. These are denoted by P and N sides, respectively, and the ‘gradients’ are often defined as the value of the corresponding parameter on the N side minus its value on the P side. The electrochemical gradient is the sum of pH gradient (concentration gradient) and membrane potential (electrical gradient). pH gradient is the (log-transformed) difference in the concentration of protons, H+, across the membrane. Protons tend to move down its concentration gradient to maximise the entropy of the whole system, and this tendency, i.e. potetial energy is the “chemical” part of the electrochemical potential. Intracellular/intra- orgenellar pH can be measured with pH-sensitive microelectrodes, nuclear magnetic resonance, or pH-sensitive fluorescent proteins (Loiselle and Casey 2010). Low-RMM weak acids and bases can equilibrate across the membrane independent of the membrane potential. -
Lecture 37 & 38: Electron Transport Chain and Oxidative Phosphorylation
CHM333 LECTURES 37 & 38: 4/27 – 29/13 SPRING 2013 Professor Christine Hrycyna Electron Transport and Oxidative Phosphorylation • Final stages of aerobic Mitochondrial Electron Transport oxidation of biomolecules in • How did we get here? eukaryotes occur in the mitochondrion • Summary of glycolysis glucose + 2 NAD+ + 2P + 2ADP 2 pyruvate + 2ATP + 2NADH + 2H+ • Reduced coenzymes NADH i → and FADH2 from: • Summary of the citric acid cycle (including pyruvate dehydrogenase) pyruvate + 4 NAD+ + FAD + GDP + 2 H 0 3 CO + 4NADH + 4H+ + P + GTP + FADH (1) Aerobic oxidation of 2 → 2 i 2 pyruvate by the citric acid cycle (2) Oxidation of fatty acids and amino acids Electron Transport Chain is the process by which NADH and FADH2 are oxidized and a proton gradient is formed. Electron Transport and Oxidative Phosphorylation Oxidative phosphorylation is the process of making ATP by using the proton gradient generated by the ETC. Respiration by mitochondria • Oxidation of substrates is coupled to the phosphorylation of ADP • Respiration (consumption of oxygen) proceeds only when ADP is present • The amount of O2 consumed depends upon the amount of ADP added Location of mitochondrial complexes • Inner mitochondrial membrane: a. Electron transport chain: oxidizes reduced coenzymes b. ATP synthase: machinery to synthesize ATP 280 CHM333 LECTURES 37 & 38: 4/27 – 29/13 SPRING 2013 Professor Christine Hrycyna Electron transport and oxidative phosphorylation capture the energy in the redox potential of NADH and FADH2 – 2 separate processes that are COUPLED to result in ATP production Extensive folding of IMM provides a large surface area on the matrix side to form lots of assemblies of proteins to maximize ATP production (1) Respiratory electron-transport chain (ETC) Series of enzyme complexes embedded in the inner mitochondrial membrane, which oxidize NADH and FADH2.