Chapter 15 Metabolism: Basic concepts and design Part Ⅰ: the specificity and catalytic power of enzymes the regulation of enzyme activity the transport of molecules and ions across membranes Part Ⅱ: to extract energy and reducing power from its environment to synthesize the building blocks of its macromolecules and then the macromolecules themselves Metabolism: a highly integrated network of chemical reactions contain many common motifs § 15.1 Metabolism is composed of many coupled, interconnecting reactions Cells transform different types of energy ¤ Phototrophs (photsynthetic organisms): light energy chemical energy ¤ Chemotrophs: Utilize chemical energy generated by phototrophs ion gradient: other types of chemical energy, nerve impulses, etc. mechanical energy: muscle contraction and movement synthesis biomolecules Metabolism(or intermediary metabolism) Energy is being extracted from fuels via a linked series chemical reactions and used it to power biosynthesis processes Two broad classes: ¤ Catabolism: fuels transform into cellular energy Fuels (carbohydrates, fats, etc) CO2+H2O+useful energy ¤ Anabolism: cellular energy to generate complex structures or energy-rich compounds Useful energy + small molecules complex molecules * Amphibolic pathways: either anabolic or catabolic is depended on the energy conditions in the cell. Metabolic pathways – many coupled, interconnecting reactions § 15.2 Adenosine triphosphate (ATP) – the universal currency of free energy in biological systems (XTP?) Carbohydrates and fats ATP 2 Phosphoanhydride bonds Mg2+ or Mn2+ -4 -2 -3 Pi: orthophosphate PPi: pyrophosphate ATP ADP (ATP-ADP cycle) the fundamental mode of energy exchange in biological systems ATP + NDP ADP + NTP (nucleoside diphosphate kinase) ATP + NMP ADP + NDP (nucleoside monophosphate kinase) ATP + AMP 2 ADP (adenylate kinase, myokinse) ¤ ATP + H2O ADP + Pi G0’= -7.3 kcal/mol ATP + H2O AMP + PPi G0’= -10.9 kcal/mol Under typical cellular condition: G = -12 kcal/mol A+BC+D G = G0’ + RT ln [C][D]/[A][B] ¤ A thermodynamically unfavorable reaction can be driven by a favorable reaction increase a factor of about 108. Keq of A B under standard condition: 1.15×10-3 Keq of A B under standard condition + ATP: 2.67×102 at pH 7, G°’= -7.3 kcal/mol Keq of A B under typical cellular condition + ATP : 7.7×105 G= -12 kcal/mol if nATP 108n ¤ ATP hydrolysis drives metabolism by shifting the equilibrium of coupling reactions chemical energy coupling agent protein conformation shift, e.g., muscle contraction the conc. of ion or molecule on the outside/inside of a cell, e.g., Na+/K+ pump What makes ATP a particular efficient phosphoryl-group donor ~ P: high energy bond The free energy of hydrolysis Three structural factors: 1. resonance stabilization, 2. electrostatic repulsion, 3. stabilization due to hydration ADP and Pi both effectively bind to water than ATP squiggle (~P) indication Phosphoryl transfer potential – an important form of cellular energy transformation An efficient carrier of phosphoryl groups High phosphoryl transfer potential compounds Creatine kinase: Creatine phosphate + ADP ATP + creatine In vertebrate muscle serves as a reservoir of high-potential phosphoryl groups C Sources of ATP (energy) during exercise § 15.3 The oxidation of carbon fuels an important source of cellular energy immediate energy donor Chemotrophs Phototrophs Free energy of oxidation of single-carbon compounds H2 11/20 施明哲主任 Two kinds of trapped energy of fuels oxidation 1. A high-energy phosphate compound GAP 1,3-BPG 3PGA acid Substrate-level phosphorylation ( p. 443) 2. Ion gradient formation Three stages of catabolism §15.4 Metabolic pathways contain many recurring motifs ¤ Activated carrier of phosphoryl groups, e.g., ATP ¤ Activated carrier of electrons for fuel oxidation e.g., nicotinamide adenine dinucleotide (NAD+) flavin adenine dinucleotide (FAD) ¤ Activated carrier of electrons for reductive biosynthesis e.g., reduced form of nicotinamide adenine dinucleotide phosphate (NADPH) NADH is used primarily for the generation of ATP ¤ Activated carrier of two-carbon fragments e.g., acetyl coenzyme A (CoA) NAD(P)+: nicotinamide adenine dinucleotide (phosphate) Niacin (vit. 3) (Fig. 15.17) ADP H+ + 2eH- (hydride ion) NADH vs. NADPH FAD: flavin adenine dinucleotide isoalloxazine ring Vit B2 riboflavin FMN 5C Coenzyme A: a carrier of acyl group thioester Transfer acetyl group is exergonic Acetyl CoA carries an activated acetyl group, just as ATP carries an activated phosphoryl group. Two key aspects of metabolism utilize activated carriers: 1. The use of specificity of enzymes to control the flow of free energy and reducing power, such as NAD(P)H, FADH2 2. The economy and elegance of metabolism underlie design Lys residue Act as coenzyme: are needed in small amounts in the diets at least 12 vitamins are needed react with hydroxyl radicals Vitamins: essential to the health of vertebrates but cannot be Nelson synthesized, so must be obtained in the diet. fat-soluble vitamins: A, D, E, K, all of which are derived from isoprene units water-soluble vitamins: C, B, biotin, folic acid, nicotinic acid,… No biological activity Nelson Regulate calcium uptake in the intestine and calcium levels in the kidney and bone Vitamin D2 (ergocalciferol): is added to milk and butter Vitamin A (retinol): the visual pigment of the vertebrate eye Nelson Cure acne and wrinkled skin p. 424 From fish liver oils, liver, eggs, whole milk, butter carrots, sweet potato, and other yellow vegetables β-carotenoids Deficiency: night blindness, dryness of the skin and eyes… Nelson Vitamin E: tocopherols: a substituted aromatic rings and a long isoprenoid side chain The aromatic ring reacts with and destroys the reactive oxygen species, protecting unsaturated fatty acids from oxidation. Tocopherols: in eggs, vegetable oils, and wheat germ Vit E deficiency: fragile erythrocytes for humans scaly skin, muscle weakness and wasting, sterility Vitamin K: Nelson active prothrombin formation Vit. K deficiency: hemorrhagic disease of the newborn in U.S.A., newborns are injected Vit. K Vit. K1 rich in green plant leaves Vit. K2 is formed by the intestinal bacteria K2: menaquinone Key reactions are reiterated throughout metabolism 15.5 p. 237 EC1 oxidoreductase ligase EC6 EC5 EC2 EC3 isomerase transferase hydrolase EC4 lyase p. 427 can proceed in either direction, depending on G and [reactants] and [products] Isomerization aconitase Glucose isomerase The addition of functional groups to double bonds or removal of groups to form double bonds — lyase aldolase enolase The commonalties in the diverse metabolic pathway (02) Metabolic processes are regulated in four principal ways 1.The amount of enzymes the rate of synthesis and degradation the rate of transcription of the genes that encoding enzymes 2. The catalytic activities of enzymes the reversible allosteric inhibition CTP inhibit asparate transcarbamoylase the reversible covalent modification myristoylation, phosphorylation, glycosylation, lipidation, methylationpalmitoylation, prenylation hormone coordination: epinephrine, insulin act through 2nd messengers farnesylation 3. The accessibility of substrates the flux of substrates among different compartments compartmentalization (synthesis/degradation) 4. The energy state Farnesyl transferase inhibitors are a new class of biologically active anticancer drugs. The exact mechanism of action of this class of agents is, however, currently unknown. The drugs inhibit farnesylation of a wide range of target proteins, including Ras. It is thought that these agents block Ras activation through inhibition of the enzyme farnesyl transferase, ultimately resulting in cell growth arrest. Ubiquitination: lysis protein Sumoylation: repress gene expression small ubiquitin-like modifer, SUMO KXE Energy charge: [ATP] + 0.5 [ADP] / {[ATP] + [ADP] + [AMP]} catabolism The pH of a cell anabolism Phosphorylation potential: [ATP] / [ADP] [Pi] ex. 11 0.9 Evolution of metabolic pathways RNA ribozymes 96 T 97T 96 C 97C 98T