Chapter 9: ATP synthase - A complex of several membrane proteins that provide a port through which protons diffuse.

This complex functions in chemiosmosis with adjacent electron transport chains, using the energy of a hydrogen ion (proton) concentration gradient to make ATP. ATP synthases are found in the inner mitochondrial membrane of eukaryotic cells and in the plasma membrane of prokaryotes. Chemiosmosis - The diffusion of ions across a selectively-permeable membrane. An energy-coupling mechanism that uses energy stored in the form of a hydrogen ion gradient across a membrane to drive cellular work, such as the synthesis of ATP. Most ATP synthesis in cells occurs by chemiosmosis, during cellular respiration. The Light reactions of photosynthesis generate energy by chemiosmosis. Chlorophyll loses an electron when energized by light. This electron travels down a photosynthetic electron transport chain ending on the high energy molecule NADPH. The electrochemical gradient generated across the thylakoid membrane drives the production of ATP by ATP Synthase. This process is known as photophosphorylation. Protons move into the thylakoid membrane at three points: On the lumen side of PS II, where water is oxidized, releasing H+. On the lumen side of the cytochrome b6-f complex as PQH2 is oxidized. PQ acquires 2 H+ from the stroma as it is reduced at the stromal side of PS II (the protons balance the two negative charges of the electrons). These protons are released as the electrons are removed at the lumenal side of the cytochrome complex. Also at the cytochrome complex, cytochrome complex, protons move into the thylakoid lumen by another mechanism called the "Q cycle". This mechanism is described in Fig. 7.29 of your text. As PQH2 is oxidized at the lumen side of the cytochrome complex, one of the two electrons goes to plastocyanin while the other goes to a different site on the stromal side of the cytochrome complex. PQ is reduced there, as well as at the stromal side of PS II, and it acquires 2 H+ from the stroma, just as it does at PS II. In this way, some of the electrons passing through the system bring two protons across the thylakoid membrane rather than just one.

For all these sites of proton movement into the lumen, the positioning of oxidation or reduction sites on PS II and the cytochrome complex is the basis of proton movement. Chemiosmosis Question: how does electron transport result in the phosphorylation of ADP, yielding ATP? Answer: Chemiosmotic hypothesis by Peter Mitchell. States that electron transport serves to generate an H+ gradient across the chloroplast thylakoid and mitochondrial inner membranes. The gradient is then used to make ATP. Protons are small but charged and don't move easily through membranes unless there is a protein channel for them. The ATP synthase complex provides such a channel in the thylakoid and mitochondrial inner membranes and uses the energy of proton passage to phosphorylate ADP. The ratio of this process is about 3 protons per ATP. The ATP synthase is a large protein complex consisting of about 24 protein subunits. It acts as a channel through which protons that are in the thylakoid lumen can escape back into the chloroplast stroma, driven by the electrochemical gradient. The passage of H+ through the ATP synthase provides energy for ATP synthesis. About 3 protons are passed for each ATP made. Evidence for chemiosmosis: ATP synthesis in the dark (Andre Jagendorf) Thylakoid membranes isolated from chloroplasts can be made to synthesize ATP in the absence of photosynthetic electron transport in the following experiment. Thylakoids are isolated from chloroplasts and suspended in a buffer at pH 4. The lumen spaces of these thylakoids gradually become pH 4 as H+ moves into them from the surrounding solution. Some of the pH 4 thylakoids are then removed from the pH 4 buffer and placed in a second beaker, also at pH 4. This beaker has phosphate and ADP in it but the thylakoids do not make ATP. Another portion of the pH 4 thylakoids are placed in a beaker having ADP and phosphate but buffered to pH 8. This time ATP is synthesized. This experiment is carried out in darkness, proving that light is

not directly required for ATP synthesis but that a proton gradient between the thylakoid lumen and the outside is required.

Uncouplers (Robert Hill) Some chemicals act to neutralize the proton gradient built up by photosynthetic electron transport, for example dinotrophenol acts as a proton carrier, shuttling protons back across the thylakoid membrane as quickly as electron transport can move them in the other direction. They inhibit ATP synthesis with affecting electron transport. Electron transport accelerates because it is no longer pumping protons against a gradient. The

fact that uncouplers can abolish ATP synthesis but have no negative effect on electron transport is more evidence in support of the chemiosmotic hypothesis. Catabolic pathway - A metabolic pathway that releases energy by breaking down complex molecules to simpler compounds. Cytochrome - An iron-containing protein that is a component of electron transport chains in the mitochondria and chloroplasts of eukaryotic cells and the plasma membranes of prokaryotic cells. Electron transport chain - A sequence of electron carrier molecules (membrane proteins) that shuttle electrons during the redox reactions that release energy used to make ATP. Oxidation - The loss of electrons from a substance involved in a redox reaction. Oxidative phosphorylation - The production of ATP using energy derived from the redox reactions of an electron transport chain; the third major stage of cellular respiration. Oxidizing agent - The electron acceptor in a redox reaction. Reducing agent - The electron donor in a redox reaction. Proton-motive force - The potential energy stored in the form of An electrochemical gradient, generated by the pumping of hydrogen ions across a biological membrane during chemiosmosis. Substrate-level phosphorylation - The formation of ATP by an enzyme directly transferring a phosphate group to ADP from an intermediate substrate in catabolism. Chapter 10: Photosynthesis: Photosynthesis consists of the light reactions (photo) and Calvin cycle (synthesis) Light = photochemical reactions (thylakoids): Split H2O, Release O2, Reduce NADP+ to NADPH, Generate ATP from ADP by photophosphorylation The Calvin cycle = photosynthetic carbon reduction cycle = PCR cycle (in the stroma)

Forms sugar from CO2, using ATP and NADPH The PCR cycle begins w/ carbon fixation, CO2 into organic molecules. Excitation of Chlorophyll by Light - Excited electrons: Fluorescence, Heat , Resonance energy transfer, Redox reactions First chemical reaction - A primary electron acceptor in the reaction center accepts an excited electron from chlorophyll a. Redox reactions in the Electron Transport Chain (ETC) During light reactions cyclic and linear routes for electron flow Linear electron flow - primary pathway - involves both photosystems, produces ATP and NADPH using light energy. A photon hits a pigment, its energy is passed among pigment molecules, excites P680. Excited electron from P680 transferred to primary electron acceptor = pheophytin (chlorophyll a w/o Mg 2+) Absorption spectrum- The range of a pigments ability to absorb various wavelengths of light; also a graph of such a range. Action spectrum - A graph that profiles the relative effectiveness of different wavelengths of radiation in driving a particular process. ATP (adenosine triphosphate) - An adenine-containing nucleoside triphosphate that releases free energy when its phosphate bonds are hydrolyzed. This energy is used to drive endergonic reactions in cells. Autotroph - An organism that obtains organic food molecules without eating other organisms or substances derived from other organisms. Autotrophs use energy from the sun or from the oxidation of inorganic substances to make organic molecules from inorganic ones. Bundle-sheath cell - in C4 plants, a type of photosynthetic cell arranged into tightly packed sheaths around the veins of A leaf. C3 plant - A plant that uses the Calvin cycle for the initial steps that incorporate CO2 into organic material, forming a three-carbon compound as the first stable intermediate.

C4 plant - A plant in which the Calvin cycle is preceded by reactions that incorporate CO2 into a four-carbon compound, the end product of which supplies CO2 for the Calvin cycle. Calvin cycle - The second of two major stages in photosynthesis (AFTER the light reactions), involving fixation of atmospheric CO2 and reduction of the fixed carbon into carbohydrate. Crassulacean acid metabolism (CAM) - An adaptation for photosynthesis in arid conditions, first discovered in the family Crassulaceae. In this process, a plant takes up CO2 and incorporates it into a variety of organic acids at night; during the day, CO2 is released from organic acids for use in the Calvin cycle. CAM plant - A plant that uses crassulacean acid metabolism, an adaptation for photosynthesis in arid conditions. In this process, carbon dioxide entering open stomata during the night is converted to organic acids, which release CO2 for the Calvin cycle during the day, when stomata are closed. Carbon fixation - The initial incorporation of carbon from CO2 into an organic compound by an autotrophic organism (a plant, another photosynthetic organism, or a chemoautotrophic prokaryote).

Carotenoid - An accessory pigment, either yellow or orange, in the chloroplasts of plants and in some prokaryotes. By absorbing wavelengths of light that chlorophyll cannot, carotenoids broaden the spectrum of colors that can drive photosynthesis. Chlorophyll - A green pigment located within the chloroplasts of plants and algae and in the membranes of certain prokaryotes. Chlorophyll a participates directly in the light reactions, which convert solar energy to chemical energy. Chlorophyll a - A photosynthetic pigment that participates directly in the light reactions, which convert solar energy to chemical energy. Chlorophyll b - An accessory photosynthetic pigment that transfers energy to chlorophyll a. Cyclic electron flow - A route of electron flow during the light reactions of photosynthesis that involves only photosystem I and that produces ATP but not NADPH or O2. Electromagnetic spectrum - The entire spectrum of electromagnetic radiation ranging in wavelength from less than a nanometer to more than a kilometer. Glyceraldehyde-3-phosphate (G3P) - A three-carbon carbohydrate that is the direct product of the Calvin cycle; it is also an intermediate in glycolysis. Heterotroph - An organism that obtains organic food molecules by eating other organisms or substances derived from them. Leaf - The main photosynthetic organ of vascular plants. Light reactions - The first of two major stages in photosynthesis (BEFORE the Calvin cycle). These reactions, which occur on the thylakoid membranes of the chloroplast or on membranes of certain prokaryotes, convert solar energy to the chemical energy of ATP and NADPH, releasing oxygen in the process. Light-harvesting complex - A complex of proteins associated with pigment molecules (including chlorophyll a, chlorophyll b, and carotenoids) that captures light energy and transfers it to reaction-center pigments in a photosystem.

Linear electron flow - A route of electron flow during the light reactions of photosynthesis that involves both photosystems (I and II) and produces ATP, NADPH, and O2. The net electron flow is from H2O to NADP+. Mesophyll - The ground tissue of a leaf, sandwiched between the upper and lower epidermis and specialized for photosynthesis.

Mesophyll cell - In C4 plants, a type of loosely arranged photosynthetic cell located between the bundle sheath and the leaf surface. NADP+ - Nicotinamide adenine dinucleotide phosphate, An electron acceptor that, as NADPH, temporarily stores energized electrons produced during the light reactions. PEP carboxylase - An enzyme that adds CO2 to phosphoenolpyruvate (PEP) to form oxaloacetate in C4 plants. It acts prior to photosynthesis. Photoautotroph - An organism that harnesses light energy to drive the synthesis of organic compounds from carbon dioxide. Photon - A quantum, or discrete quantity, of light energy that behaves as if it were a particle. Photophosphorylation - The process of generating ATP from ADP and phosphate by means of a proton-motive force generated across the thylakoid membrane of the chloroplast or the membrane of certain prokaryotes during the light reactions of photosynthesis. Photosynthesis - The conversion of light energy to chemical energy that is stored in sugars or other organic compounds; occurs in plants, algae, and certain prokaryotes. Photosystem - A light-capturing unit located in the thylakoid membrane of the chloroplast or in the membrane of some prokaryotes, consisting of a reaction-center complex surrounded by numerous light-harvesting complexes. There are two types of photosystems, I and II; they absorb light best at different wavelengths. Photosystem I (PS I) - One of two light-capturing units in a chloroplasts thylakoid membrane or in the membrane of some prokaryotes; it has two molecules of P700 chlorophyll a at its reaction center. Photosystem II (PS II) - One of two light-capturing units in a chloroplasts thylakoid membrane or in the membrane of some prokaryotes; it has two molecules of P680 chlorophyll a at its reaction center. Primary electron acceptor - In the thylakoid membrane of a chloroplast or in the membrane of some prokaryotes, a specialized molecule that shares

the reaction-center complex with a pair of chlorophyll a molecules and that accepts an electron from them. Producer - An organism that produces organic compounds from CO2 by harnessing light energy (in photosynthesis) or by oxidizing inorganic chemicals (in chemosynthetic reactions carried out by some prokaryotes). Reaction-center complex - A complex of proteins associated with a special pair of chlorophyll a molecules and a primary electron acceptor. Located centrally in a photosystem, this complex triggers the light reactions of photosynthesis. Excited by light energy, the pair of chlorophylls donates an electron to the primary electron acceptor, which passes an electron to an electron transport chain. Rubisco - Ribulose bisphosphate (RuBP) carboxylase, the enzyme that catalyzes the first step of the Calvin cycle (the addition of CO2 to RuBP). Spectrophotometer - An instrument that measures the proportions of light of different wavelengths absorbed and transmitted by a pigment solution. Stoma - (plural, stomata) A microscopic pore surrounded by guard cells in the epidermis of leaves and stems that allows gas exchange between the environment and the interior of the plant. Stroma - Within the chloroplast, the dense fluid of the chloroplast surrounding the thylakoid membrane; involved in the synthesis of organic molecules from carbon dioxide and water. Thylakoid - A flattened membranous sac inside a chloroplast. Thylakoids exist in an interconnected system in the chloroplast and contain the molecular machinery used to convert light energy to chemical energy. Visible light - That portion of the electromagnetic spectrum that can be detected as various colors by the human eye, ranging in wavelength from about 380 nm to about 750 nm. Wavelength - The distance between crests of waves, such as those of the electromagnetic spectrum.