NOTES. CARBOHYDRATE METABOLISM CITRIC ACID CYCLE (KREBS CYCLE) RAs Gee oc} + Generates energy in the form of GTP, NADH, and FADH, + Occurs in mitochondria * Starts with acetyl-CoA — CO, Process + Acetyl-CoA + oxaloacetate (via citrate synthase) — citrate + CoA * Citrate (via aconitase) — isocitrate + Isocitrate + NAD* (via isocitrate dehydrogenase) — a-ketoglutarate + NADH + CO, + Rate-limiting step + a-ketoglutarate + NAD+ + CoA-SH (c-ketogiutarate dehydrogenase) succinyCo + NADH + Ci + Requires five cofactors: thiamine, lipoic acid, CoA, FAD, NAD* =» See mnemonic + Succinyl-CoA + phosphate + GDP (via succinate thiokinase) — succinate + GTP + Succinate + FAD* (via succinate dehydrogenase) — fumarate + FADH; + Fumarate + H,0 (via fumarase) + malate + Malate + NAD* (via malate dehydrogenase) = oxaloacetate + NADH * Oxaloacetate then enters next cycle + Generates one GTP molecule, three NA\ molecules, one FADH, molecule \_) MNEMONIC: T-rex Loves ~ and Cares For Nechos Five required cofactors Thiemine Upoic acid CoA FAD" NAD Te T-rex Loves & Cares For Nachos Figure 14 Mnemonic for the five cofactors required by a-ketoglutarate dehydrogenase. OSMOSISORG 12 OSMOSIS.ORG teracigcon Shy ca CITRATE a ‘ -° o-_ACONTASE caveare syurwase AR Sea OXALOACE TATE aL. JL. ISOCITRATE ISOCTTRATE DEHYDROGENASE saver Naps Nabi co, A mney. Matar 3 e op OCKETOGLUTARATE It cain) F ‘ KETOGLUTARATE Fumanae wvorase L—H,0 Naw ‘DEHYDROGENASE ° co, FUMARATE ee febrero SUCCINYL-CoA T hen! FAD, con-SA. swcenareponrocowse Sf SUCCINATE THOKINASE, “ ‘SUCCINATE 3 NADH—> 9 ATP. LOMTRIC aciD cyete 1 FADK,——> 2 ar. 12 ATP 16%? 11? Figure 1.2 The citric acid (Krebs) cycle. Each acetyl-CoA molecule generates 12 ATP, ELECTRON TRANSPORT CHAIN & OXIDATIVE PHOSPHORYLATION xidativ OU eae ut osphorylation Oxidative phosphorylation + FADH, from cytoplasm comes through + Generates energy as ATP * Occurs in inner mitochondrial membrane Electron transport chain * Series of proteins, ipids, metals that faciltates electron movernent — proton gradient used to create ATP + Starts with electron donors NADH, FADH, = NADH from cytoplasm comes through malate-espartete shuttle Glycerol-3-phosphate shuttie » NADH donates electron to complex | (contains flavin mononucleotide, iron- sulfur centers) > NAD* = FADH, donates electron to complex It (le. succinate dehydrogenase) > FAD lectrans from either complex flow into coenzyme Q (ubiquinone) = Coenzyme Q passes electrons to ‘cytochromes [proteins with hemegroups — Fe + ++ Fe%): complex ill [cytochromes b and cl) + cytochrome c— complex IV (cytochrome oxidase: cytochromes a, a3) + oxygen + Movement of electrons — electrical current = complexes | I IV use this energy to pump protons across inner mitochondrial membrane ‘COENZYMI Chapter 1 Biochemistry: Carbohydrate Metabolism + Protons can move back into mitochondria ‘through F, ~ proton gradient forms, powering F: ADP + ATP + Collectively called complex V + AN ADPIATP antiport pumps ATP into cytoplasm of the cell, supplies complex V with new ADP ae COMPLEX I!) [etschrome o J [COMPLEX IV} = 2 Ho Figure 1.3 The flow of electrons through the electron transport chain, which takes place in the inner mitochondrial membrane. OsMOSISORG 3OSMOSIS.ORG Figure 1.4 Oxidative phosphorylation. The passing of electrons along the electron transport chain generates an electrical current, which provides the eneray that allows complexes |, Il, and IV to pump protons into the space between the inner and outer mitochondrial membranes. This ‘creates a gradient across the inner mitochondrial membrane. The protons use proton chennel F, to flow down the gradient, back into the mitochondrial matrix. F, is attached to enzyme F,, an ATP synthase, which uses the proton gradient to phosphorylate ADP — ATP. GLUCONENGENESIS + Synthesis of glucose from non- carbohydrate substrates © E.g. amino acids, lactate, alyceral + Occurs primarily in liver cells; also in epithelial cells of kidney, intestine * Inside cytoplasm, mitachondria, endoplasmic reticulum + Starts with glycogenolysis after glucose depletion Process + Like backwards glycolysis, with three exceptions + Obtaining pyruvate = Lactate (via lactate dehycrogenase) —+ pyruvate = Amino acids (nat leucine, lysine): 2.4 alanine (via alanine transaminase) — pyruvate * Obtaining ATP, glycerol Triacylglyceride breakdown — fatty acids and glycerol — acetyi Cod + ATP (B-oxidation) + Pyruvate (vis pyruvate carboxylase) oxaloacetate + Oxaloacetate (malate dehydrogenase) malate + Malate leaves mitochondria: malate (vis malate dehydrogenase) — oxaloacetate + Oxaloacetate (via PEPCK) — phosphoenoloyruvate (PEP) + PEP undergoes reversed glycolysis reactions until dihydroacetone-phasphate (Oar) + Alternatively, glycerol (via glycerol kinase) — glycerol-3-phosphate;Chapter 1 Biochemistry: Carbohydrate Metabolism alycerol-3-phosphate (via alycerol-3- + Fructose-6-phosphate (via isomerase) + phosphate dehydrogenase) — DHAP glucose-6-phosphate + DHAP (via aldolase) — fructose-1,6- + Glucose-6-phosphate (via glucose-6- bisphosphate phosphatase) — glucose + Fructose-1,6-bisphosphatase — fructose- 6-phosphate = Rate-limiting step =| a oes o ll oom mcm SE ae WM on anne «a 2 Figure 1.5 The process of gluconeogenesis, OSMOSISORG 56 OSMOSIS.ORG GLYCOGEN METABOLISM + Polymer of glucose molecules linked by aiycosidlc bonds + Stores energy in skeletal muscle, liver Glycogen synthesis + Glucose + phosphate (via hexokinase) + glucose-6 phosphate + Glucose-6 phosphate (via phosphoglucomutase] —+ glucose-1- phosphate + energy (UTP) + Glucose-1-phosphate + UTP (via UDP- Glucose pyrophosphorylase) — UDP- glucose + UDP-glucose added (vie glycogen synthase) to glycogen branchiaiycogenin (= alpha-1,4-glycosiaic bond) + Branching enzyme cuts off part of glucose chain, creates branch (— alpha-1,6- glycosidic bond) Glycogen breakdown, AKA glycogenolysis + Glucagon — liver breakdown of glycogen * Epinephrine — skeletal muscle breakdown of glycogen + Glycogen phosphorylase cleaves aloha-1,4 bonds on branches; catalyzes phosphate transfer to alucose residue — one glucose- L-phosphate is released at a time += Repeats until branch is only 4 glucose units long + Debranching enzyme: 4-alpha- glucanotransferase moves 3 glucose units ff branch, onto main chain; alpha-1,6- glucosidase cleaves last remaining glucose + Cleaved glucose-1-phosphate (via phosphoglucomutase) — glucose-6- Phosphate = With glucose-6-phosphate + In liver cells, glucose-6-phosphatase removes phosphate — free glucose into blood + In skeletal muscle, glucose-G-phosphate glycolysis pathway Regulation + Principles = Glycogen synthase: active without phosphate = Glycogen phosphorylase: active with phosphate + Hormones = Insulin: binds to membrane tyrosine kinase receptors — protein phosphatase removes phosphates — glycogen synthase activates, glycogen phosphorylase deactivates * Glucagon: binds to membrane G-protein ‘coupled receptors (in liver) +» ATP (adeny¥yl cyclase) + cAMP — kinase A adds phosphates — alycogen phosphorylase activates, glycogen synthase deactivatesChapter 1 Biochemistry: Carbohydrate Metabolism GLYCOGEN SYNTHESIS GLUCOSE-1 PHOSPHATE vr pP-cLucose, step a fend of GLYCOGEN BRANCH/ NEW LINEAR GLYCOGEN MOLECULE \aroua-tsreauveasiore Bono Sih RE OOS UDP-GLUCOSE. STEPS ALPHA 6 GuyCosioic BoD ——> [BRANCHED GLYCOGEN TREE. Figure 1.6 The process of glycogen synthesis. OSMOSISORG 7GLYCOGEN BREAKDOWN ‘STEP1 t GLYCOGEN Socoboo CORSO oo * raved ctvcocen yp OE, & & o civeese- mospuare STEP ae DEBRANCHING ENZYME: 2222S @eoeocoocsoo STEP ab @0250555.5-5. samemaoame alpha .6-glucesidae Gy ALUCOSE STEPS , : ] se oho = & %& aves sane sucoseenespune MUSCLES NO GLUCOSE-6-PHOSPHATASE 1 USES GLUCOSE-6-PHOSPHATE Leuvcoursis prruuay Guakes enerey Figure 17 Glycogen breakdown. The process is completed cifferently in the liver and skeletal muscles due to the respective presence and absence of glucose-6-phasphatase in each. 8 — OSMOSIS.ORGREGULATION ALINSULIN ‘TYROSINE KINASE. RECEPTOR PROTEIN ‘sailed REMOVES PHOSPHATES GLYCOGEN GLYCOGEN SYNTHASE. PHOSPHORYLASE MAKES ACTIVE MAKES INACTIVE. ToLycocen FGLYCOGEN SYNTHESIS BREAKDOWN Figure 1.8 The role of insulin in the regulation of glycogen levels. Chapter 1 Biochemistry: Carbohydrate Metabolism REGULATION 2. GLUCAGON G-PROTEIN COUPLED RECEPTOR 4 ADENYLYL CYCLASE Gare — cAMP ae KINASE AL 4 ADDS PHOSPHATES GLYCOGEN GLYCOGEN ‘SYNTHASE PHOSPHORYLASE MAKES INACTIVE MAKES ACTIVE | SYNTHESIS BREAKDOWN Figure 1.9 The role of glucagon in the regulation of glycogen levels. GLYCOLYSIS + Energy-producing breakdown of glucose into pyruvate * Oceurs in cytoplasm of all cells PROCESS * Glucose transporter (GLUT) carries glucose into cell + Kinases (hexokinase, glucokinase) phosphorylate glucose — conformational change, ie, glucose can‘ dfuse out) glucase-6-phosphate 2 Uses one ATP molecule + Glucose-6-phosphate (via phosphoglucoisomerase) — fructose-6- phosphate + Fructose-6-phosphate (Via phosphofructokinase-1) — fructose-1,6- bisphosphate = Rate-limiting step 2 Uses one ATP molecule OSMOSISORG 910 OSMOSISORG Enzyme activation + Fructose-6-phosphate (via phosphofructokinase-2) — fructose-2.6- bisphosphate + Up-regulated by insulin; down- regulated by glucagon © Fructose-2,6-bisphosphate activates phosphofructokinase-1 + Fructose-1,6-bisphosphate (via aldolase) = glyceraldehyde 3-phosphate (63°) + dinydroacetone-phosphate (DHAP) = DHAP (via isomerase) — G3P + 2x GP molecules per glucose * G3P [via G2P-dehydrogenase) + 1,3-diphosphoglycerate (1.3-BPG); H- + NAD*— NADH (x2] + 2x NADH enter electron transport chain + 1,3-8PG + ADP (via ohosphoalycerate kinase) — 3-phosphoglycerate + ATP (<2) += Creates two ATP molecules + 3:phosphoalycerate (via mutase) —+ 2-phosphogiycerate (x2) + 2:phosphoglycerate (via enolase) > phosphoenalpyruvate (PEP) + H.0 (2) + PER + ADP (via pyruvate kinase) + pyruvate + ATP (2) = Creates two ATP molecules + Up-regulated by fructose-1,6- bisphosphate (feed-forward regulation) = Down-regulated by ATP, alanine + In total, process generates two ATP molecules + In cells with oxygen, pyruvate enters citric acid cycle, electron transport chain to make more ATP + 20-32 in total PENTOSE PHOSPHATE PATHWAY + Synthesis of ribose, NADPH trom unused glucose * Occurs in cytoplasm of all cells Irreversible oxidative phase + Glucose-6-phosphate + NADP (via glucose-6-phosphate dehydrogenase) 6-phosphogluconate + NADPH += Rate-limiting step + @-phosphogluconate + NADP* (6-phosphogluconate dehydrogenase) = ripulose-8-phosphate + NADPH + CO, Reversible non-oxidative phase + Two options: += Ribulose-5-phosphate (via isomerase} — ribose-5-phosphate » Ribulose-5-phosphate = xylulose-5-phosphate ja epimerase)Chapter 1 Biochemistry: Carbohydrate Metabolism G ercray (are) Tracker Ao spec acre in is ere et a \ Renae 0. Co dhe. crn sam eG Oe EO coal socanef manne ngs Lng sal ne (on Lag J) roense [fore x2 Perse ONO + MAOH x2 —> erecrRON TRansPoRT cHaIN 0 227 DIt merci 2xATP. 0 some x2 tt wuTase, 02, a-rnosrioaticenare x2 se Wie -o PEP, 4 no nein are, o. . sone 2 . ATP x2 ae rumen A MITOCHONDRION cmc aio (ness) os Bourn 30-82 ATP Figure 140 Glycolysis. OSMOSISORG 11Figure 141 Pentose phosphate pathway. 12 OSMOSISORG