A. Mitochondrial structure and function Localization of metabolic functions within the mitochondrion 2. Molecular basis of oxidative  phosphorylation A. Molecular basis of oxidation: Electron-  transport chain B. Molecular basis of phosphorylation: ATP synthase Direct experimental evidence supporting the rotational catalysis. The ATP synthase is a reversible coupling device  Other roles for the proton-motive force in addition to ATP synthase C. Mithchell’s Chemiosmotic theory (1961) 3. Chloroplast and photosynthesis A. Comparison of a mitochondrion and a chloroplast.  B. Photosynthesis 4. Organelle DNA and protein importing A. Organelle DNA C. The transport protein into Mit. And Chl. Light-dependent reaction: Electron transport in the thylakoid membrane and noncyclic photophosphorylation: Cyclic photophosphorylation: Changes in redox potential during photosynthesis. Carbon dioxide fixation and the synthesis of carbohydrate in C3 plants (Calvin cycle) Figure 14-43 The initial reaction in carbon fixation. This reaction, in which carbon dioxide is converted into organic carbon, is catalyzed in the chloroplast stroma by the abundant enzyme ribulose bisphosphate carboxylase. The product, 3-phosphoglycerate, is also an important intermediate in glycolysis: the two carbon atoms shaded in blue are used to produce phosphoglycolate when the enzyme adds oxygen instead of CO2。The structure and function in C4 plants The size range of organelle DNA is similar to that of viral DNAs. Mit DNA: from <6,000bp (plasmodium falciparum) ～>300,000bp (some land plants).  DNA of Mit genome (in mammals) ≈16,500bp(<0.001% of nuclear genome) ; Chl genomes are about 10 times larger and contain about 120 genes. Chl DNA: from 70,000 to 200,000bp (genome of land plants); Genes in mtDNA encode rRNAs, tRNAs, and some mitochondrial proteins Human mt DNA: 16,569bp  2 rRNAs, 22 tRNAs, 13 polypeptides: NADH reductase. 7 sub. Cty b-c1 complex. 1 cytb Cyt oxidase. 3 subunits ATP synthase: 2 F0 sub  Products of mt genes are not exported The organization of the liverwort( 地钱) Chl genome B. Mit and Chl have their own genetic systems Mit and Chl are organelles semiautocephaly.  The synthesis of mt proteins is coordinated * * Energy Generation in Mitochondria and Chloroplasts Chapter 7 (1)  Mitochondria: in all eukaryotic cells The relationship between the structure and function of mit. (2) Chloroplasts: in plant cells The relationship between the structure and function of chl. Mit: Oxidative phosphorylation →ATP Chl: Photosynthesis →ATP + NADPH →Sugar The size and number of mitochondria reflect the energy requirements of the cell. 1. Mitochondria and oxidative phosphorylation Figure 7-4 Relationship between mitochondria and microtubules. Figure 7-3 Mitochondrial plasticity. Rapid changes of shape are observed when a mitochondrion is visualized in a living cell.  Figure 7-5 Localization of mitochondria near sites of high ATP utilization in cardiac muscle and a sperm tail. Inner and outer mitochondrial membranes enclose two spaces: the matrix and intermembrane space. Outer membrane: Contains channel-forming protein, called Porin. Permeable to all molecules of 5000 daltons or less. Inner membrane (Impermeability): Contains proteins with three types of functions: (1) Electron-transport chain: Carry out oxidation reactions; (2) ATP synthase: Makes ATP in the matrix; (3) Transport proteins: Allow the passage of metabolites Intermembrane space:  Contains several enzymes use ATP to phosphorylate other nucleotides. Matrix: Enzymes; Mit DNA, Ribosomes, etc.  Figure 14-6 Fractionation of purified mitochondria into separate components. These techniques have made it possible to study the different proteins in each mitochondrial compartment. The method shown, which allows the processing of large numbers of mitochondria at the same time, takes advantage of the fact that in media of low osmotic strength water flows into mitochondria and greatly expands the matrix space (yellow). While the cristae of the inner membrane allow it to unfold to accommodate the expansion, the outer membranewhich has no folds to begin withbreaks, releasing a structure composed of only the inner membrane and the matrix. B. Specific functions localized within the Mit by disruption of the organelle and fractionation Outer membrane: Phospholipid synthesis fatty acid desaturation Fatty acid elongation Inner membrane: Electron transport Oxidative phosphorylation Metabolite transport Intermembrane space Nucleotide phosphorylation Matrix Pyruvate oxidation TCA cycle  oxidation of fats DNA replication, RNA transcription, Protein translation The structure of the ATP synthase F1 particle is the catalytic subunit; The F0 particle attaches to F1 and is embedded in the inner membrane. F1: 5 subunits in the ratio 3 :3 :1 :1 :1 F0: 1a：2b：12c F1 particles have ATP synthase activity Proton translocation through F0 drives ATP synthesis by F1: Binding Change Model and rotational catalysis Boyer proposed in 1979, and was greatly stimulated by the publication in 1994 of the structure for F1 complex (X-ray) from bovine heart mitochondria Japan researcher, Nature 386: 300, 1997. The pH and electrical gradient resulting from transport of protons links oxidation to phosphorylation. When electrons are passed to carriers only able to accept electrons, the H+ is translocated across the inner membrane. More than 2 1026 molecules (>160kg) of ATP per day in our bodies. Electrons pass from NADH or FADH2 to O2, the terminal electron acceptor, through a chain of carriers in the inner membrane (FMN, Fe-S center, Heme group Fe, CoQ); As electrons move through the electron-transport chain, H+ are pumped out across the inner membrane, and form Proton motive force; Electrons move through the inner membrane via a series of carriers of decreasing redox potential If not all the detergent is removed, what will happen?  Figure 7-26 An experiment demonstrating that the ATP synthase is driven by proton flow. By combining a light-driven bacterial proton pump (bacteriorhodopsin), an ATP synthase purified from ox heart mitochondria, and phospholipids, vesicles were produced that synthesized ATP in response to light. Summary of the major activities during aerobic respiration in a mitochondrion NADH O2: 3ATP/2e; FADH2 O2 : 2ATP/2e  生物氧化产生ATP 的统计 一个葡萄糖分子经过细胞呼吸全过程产生多少ATP？  糖酵解：底物水平磷酸化产生4 ATP（细胞质） 己糖分子活化消耗2 ATP（细胞质） 产生2 NADH，经电子传递产生4或6 ATP （线粒体）净积累6或8 ATP   丙酮酸氧化脱羧：产生2 NADH（线粒体），生成6 ATP  三羧酸循环：底物水平的磷酸化产生（线粒体）2 ATP； 产生6 NADH（线粒体），生成18 ATP； 产生2 FADH2（线粒体），生成4 ATP  总计生成36或38 ATP  Figure 14-39 The chloroplast. This photosynthetic organelle contains three distinct membranes (the outer membrane, the inner membrane, and the thylakoid membrane) that define three separate internal compartments (the intermembrane space, the stroma, and the thylakoid space). The thylakoid membrane contains all of the energy-generating systems of the chloroplast. In electron micrographs this membrane appears to be broken up into separate units that enclose individual flattened vesicles (see Figure 14-40), but these are probably joined into a single, highly folded membrane in each chloroplast. As indicated, the individual thylakoids are interconnected, and they tend to stack to form aggregates called grana.  (A) A wheat leaf cell in which a thin rim of cytoplasm containing chloroplasts surrounds a large vacuole. (B) A thin section of a single chloroplast, showing the starch granules and lipid droplets that have accumulated in the stroma as a result of the biosyntheses occurring there. (C) A high-magnification view of a granum, showing its stacked thylakoid membrane. (Courtesy of K. Plaskitt.)   Figure 7-42 Photosynthesis in a chloroplast. Water is oxidized and oxygen is released in the photosynthetic electron-transfer reactions, while carbon dioxide is assimilated (fixed) to produce carbohydrate in the carbon-fixation reactions. C. The antenna complex and photochemical reaction center in a photosystem