Biochemical Seminar I PDF

OUR LADY OF FATIMA UNIVERSITY COLLEGE OF PHARMACY PHARMACEUTICAL SEMINAR I TOPIC OUTLINE NO true Nucleus TRUE NUCLEUS (1) Basic Concepts of Biochemistry...

OUR LADY OF FATIMA UNIVERSITY COLLEGE OF PHARMACY PHARMACEUTICAL SEMINAR I TOPIC OUTLINE NO true Nucleus TRUE NUCLEUS (1) Basic Concepts of Biochemistry NO membrane bound organelles Mitochondria, Lysosomes, ER, (2) Biomolecules Golgi apparatus  Carbohydrates 30s + 50s = 70s 40s + 60s = 80s  Lipids PROTOZOA → bacteria Plants, Animals, Fungi  Nucleic acids  Proteins PARTS OF THE CELL Biosynthesis Functions Classification Physilogical significance (3) Metabolic Pathway BIOCHEMISTRY  study of chemistry of life  the science concerned with the chemical basis of life  chemical constituents of living cells and with the reactions and processes they undergo  a study of the compounds and processes associated with living organisms  the study of the chemical substances found in living organisms and the chemical interactions of these substances with each other  “BIOS” - life IMPORTANT INFO Any hindrances or blockage in biochemical events will manifest a disease. CELL MEMBRANE  AKA “PLASMA MEMBRANE/ PLASMA LEMMA” CELL  The outermost structure of the cell that decides its contour is  Basic unit of life the cell membrane.  Aggregation of cells → Tissues → Organs → Organ system →  SEMIPERMEABLE, allows only non-polar molecules to enter the Organism cell  Molecular Composition  Components: → Water (70-75%) → Phospholipids → Organic Molecules/Biomolecules (essential to life) - main component of lipid bilayer complecx  Nucleic Acids - 1 hydrophobic head and 2 hydrophilic tail  Proteins → Glycoprotein and glycolipids  Carbohydrates  Glycoprotein  Lipids - is a conjugate protein where the prosthetic group is a carbohydrate Biomolecule Building blocks Major function - part of the integral membrane of the proteins - play a vital role in cell-cell interaction and Nucleic acids Deoxyribonucleic acid Genetic material - DNA Ribonucleic acid template for protein pathogenic response bu the host - RNA synthesis  Glycolipids - lipids with carbohydrate attached to a glycosidic Proteins α-amino acid Cell molecules that covalent bond carry out work (ex. - maintain the stability of cell membrane and enzyme) facilitate cellular recognition, which is crucial to the Carbohydrates Monosaccharides Short term of energy immune response as (glycogen is the → Cholesterol main storage form of - biosynthesize by all animal cells glucose) - essential structural component of animal cell Lipids Fatty acids Numerous functions membrane (ex. Membrane - yellowish crystalline solid component of the long - precursor in the biosynthesis of steroid term storage of energy hormones, bile acids, and Vitamin D as fat) - fluidity and consistency of cell Main way of the body → Arachidonic acid to store excess - polyunsaturated omega-6 fatty acid (20:4) with a nutrient double bond at 5, 8, 11, and 14 covalently bond to esterified form in the cell membrane of most of the → Inorganic Molecules body cells  Trace Elements - irritation and injury, the arachidonic acid releases - ex. Na, K, Mg, Cu, Fe, and Anions (Chloride, and oxygenates by the enzyme system leading to Phosphate, Bicarbonate, Sulfate, Iodide, and the formation of important inflammatory mediator Fluoride) called EICOSANOIDS - a substrate for eicosanoid synthesis, from COX CELL pathway to Prostaglandins and TX or PROKARYOTE EUKARYOTE thromboxanes; LOX pathway will form LTs or 1  OUR LADY OF FATIMA UNIVERSITY COLLEGE OF PHARMACY PHARMACEUTICAL SEMINAR I leukotrienes; and CYP 450 pathway form various  HEMOPHILIA B epoxy, hydroxy, and dihydroxy derivatives - deficiency of factor IX IMPORTANT INFO → Nuclear Membrane Fluid Mosaic Model - protects the genetic material - describes the structure of cell membrane  Nuclear pores - indicates that the cell is not solid and it is flexible with - for the exchange of material between nucleus similar consistency to a vegetable oil, so that all individual and the rest of the cell molecule are all floating in a fluid medium and capable of  Nuclear envelope moving side-ways within the cell membrane - two layered outer limit that separates from cytoplasm Organism Membrane/wall Sterol present  Cytoplasm Plants Cellulose Phytosterol and B- - numerous enzymes, proteins, and many other Sitosterol solutes are found in the cytosol or liquid portion of Bacteria Peptidoglycan the cytoplasm, which is the main site for glycolysis, Fungi Chitin Ergosterol HMP shunt, and activation of amino acid and fatty acid synthesis  Cell division IMPORTANT INFO - process by which a parenteral cell divides in two Cellulose a polysaccharide consisting of a linear chain of or more daughter cells several hundreds to many thousand link dextrose glucose  Centromere units; important structural components of the primary cell - a constricted region of chromosome that separate wall of green plants it into short r (p) and long arm (q) PHYTOSTEROL is similar to cholesterol and serve as Characteristics Prokaryote Eukaryote structural component of biological membrane of plants Chromosome Single circular Paired linear BETA-SITOSTEROL a white waxy powder with Chromosome Nucleoid Nucleus characteristic odor and component of food additive E499 location Nucleolus Absent Present PEPTIDOGLYCAN or murine; a polymer consisting of Extra chromosomal Plasmid Mitochindria and sugar and amino acid that forms a mesh-like-layer DNA Chloroplast outside the plasma membrane of most bacteria forming Site of cellular Cell membrane Mitochondria the cell wall respiration Ribosomes 30s and 50s/70s 40s and 60s/80s in CHITIN (C8H13O5N) a long-chain polymer of N- cytoplasm/70s in acetylglucosamine; an amide derivative of glucose organelles Locomotion Rotating flagella and Undulating flagella gliding and cilia, and also NUCLEUS amoeboid  “CONTROL center of the cell” Pili Sex or attachment Absent  information center of eukaryotic cell. It is mainly concentrated in pill the form of CHROMOSOMES  Components: IMPORTANT INFO → Nucleolus PLASMID is a small-circular double stranded DNA for - site of ribosomal assembly or ribosoma; RNA antibiotic resistance synthesis; a nuclear matrix - contains enzymes involve in the synthesis of MITOCHONDRIA is the power-house of the cell or DNA and RNA electron transport enzyme fro citric acid cycle , beta - dark spot in the middle of the nucleus that helps oxidation, and respiratory chian make ribosomes → Chromosomes PILI adds up to the virulence of bacteria or form of  Chromatin attachment - a thread-like loose network; not present in cell division ORGANELLES  Chromosomes - a rod-like coiled network; present in cell division  a specialized sub-unit within a cell that has a specific function - found in the nucleus  Components: - made up of DNA and proteins → Mitochondria - XX (23 female chromosomes) - It has an inner and outer membrane which - XY (23 male chromosomes) consists of proteins and phospholipids - TRISOMY 21 OR DOWN SYNDROME term for - “Powerhouse of the cell” due to production of having an extra chromosomes ATP/energy - Y LINKED a condition if the mutated gene that - CARDIOLIPIN an inner phospholipid present causes the disorder is located at the Y and an important component of inner chromosome; father to son disorder mitochoindrial membrane, which constitute 20% of - HEMOPHILIA is the lack of clotting factor which the total lipid composition produces excessive bleeding - CRISTAE a term for inner division  HEMOPHILIA A → Lysosomes - deficiency of factor VIII - Called as ‘SUICIDAL BAGS’ or ‘DEMOLITION - most common SITE; of the cell 2  OUR LADY OF FATIMA UNIVERSITY COLLEGE OF PHARMACY PHARMACEUTICAL SEMINAR I - Lysosomal enzymes digest the molecules Scrapie Sheep brought into the cell by phagocytosis Creutzfeldt-Jakob disease Humans - breakdown excess or worn out cell parts to destroy viruses and bacteria → Vacuole - 3 types of cell death → Plastids 1. Necrosis → Cell wall - aka “cell murder” - the cell undergo necrotic death if Animals Plant cells the cell membrane is damage or due Shape Round or irregular Rectangular to the decrease oxygen supply and if shape energy production is block Cell wall Absent (Cell Present 2. Atropy membrane) - due to absence of survival factors Plastids None Present 3. Apoptosis Vacuole 1 or more 1 - a program cell death - responsible for metastasis → Peroxisomes CELL FUNCTION - aka Peroxidases and Catalases  Transport - are also small vesicles surrounded by a → Diffusion membrane. They are also called as - a passive process of transport MICROBODIES - a single substance move from area of high conc to are - they contain enzymes for H2O2 metabolism of low conc - ZELLWEGER’S SYNDROME/SPECTRUM  Passive DISORDERS  Simple - a defective in gene required in the formation - (like dissolves like) and function of peroxisomes  Facilitated - characterized by a low muscle tone - saturable and limited by numbers of carriers (hypotonia), development delay, early death, - w/ a carrier of transport protein seizure, and skull malformations  Active - an adrenoleukodystrophy damage, a white - moves from area of low conc to area of high matter of the brain and impairs the adrenal conc with the requirement of energy glands; children may show delayed → Endocytosis development but not have vision and - movement into the cell hearing problems until adulthood  Phagocytosis → Ribosomes - “cell eating” - very small macromolecular complexes  Pinovytosis composed of rRNA and proteins responsible in - “cell drinking” protein synthesis (translation) on mRNA templates - e.g. fats, glycerin, starch, parasite eggs, vitamins → Endoplasmic Reticulum A,B, E and K, plastic particles, hairs, yeast, ferritin, - system of fluid filled with cisterns or foldings and insulin - ROUGH E.R → Exocytosis - are involved in protein synthesis - movement out of the cell - ribosomes are present - SMOOTH E.R BIOMOLECULES - Lipids synthesis - Ribosomes are NOT present Two types of Biochemical Substances → Golgi apparatus Bioinorganic Bioorganic - are well developed in cells, which are involved in Water Carbohydrates SECRETION involved in glycosylation and Inorganic salts Lipids sulfation of proteins Proteins - modify and package protein Nucleic acid - stack of flatten socks → Plasmids Biomolecule Building blocks Bonds Examples - advantages on the host: Carbohydrates Monosaccharides Glycosidic Starch and - confer resistance to toxins and antibiotics glycogen - some encode for proteins which confer Lipids Fatty acids Ester Triglycerides virulence factors on the host and Waxes - e.g. E.coli plasmid Ent P307 codes for an Nucleic acids Nucleotides Phosphodiester DNA and enterotoxin which makes E.coli pathogenic RNA - conjugative plasmids Proteins Amino acids Peptide Pepsin and - these allow exchange of DNA between Hbg bacterial cells → Prions - infectious agents consisting of proteins but no CARBOHYDRATES nucleic acid  “Hydrates of carbon” - from proteinaceous and infectious  General formula: CnH2nOn DISEASES  Either a ketone or aldehyde and composed of Carbon, Hydrogen, Bovine spongiform Cattle and Oxygen encephalopathy (BSE)/Mad  First product of photosynthesis cow disease 3  OUR LADY OF FATIMA UNIVERSITY COLLEGE OF PHARMACY PHARMACEUTICAL SEMINAR I  Polyhydroxy compounds with aldehyde, ketone or compounds POP QUIZ that produce such substances upon hydrolysis  Source of Energy: → 1g Lipids: 9 kcal → 1g Carbo: 4 kcal → 1g Protein: 4 kcal  Functions: → Storage form of fuel → Structural elements → Proteoglycans → Glycosaminoglycans also known as mucopolysaccharide → Lubricant of skeletal joints → Major component of bacterial cell walls and soft cell coats in animal tissues → Provide adhesion between cells - e.g. Hormonal activation of cells, immune system mediation through cytokines → Transport functions (biochemical transport) e.g. Transferrin for Fe transport, Ceruloplasmin for Cu Transport → Confer biologic specificity on animal cell surface - Different blood types based on sugar moieties present/absent → Supply precursors for synthesis of vital biologic substances - Enzymes - Hormones - Precursors of DNA, RNA and ascorbic acid B. CLASSIFICATIONS OF CARBOHYDRATES  General properties: → Physical properties CARBOHYDRATES - The mono and disaccharides are white crystalline substances and sweet - Starches are amorphous powder and tasteless - The most complex cellulose is fibrous and tasteless → Chemical properties CRYSTALLINE FIBROUS - Reducing power - all mono and disaccharide containing the potentially free aldehyde or ketone group possess reducing properties MONOSACCHARIDE OLIGOSACCHARIDE A. BIOSYNTHESIS OF CARBOHYDRATES DISACCHARIDE POLYSACCHARIDE B.1. MONOSACCHARIDES  The simplest sugar; building block of carbohydrates  Contains single polyhydroxy aldehyde and ketone; contains 3-10 carbon atoms  Can be classified according to: → the number of carbon atoms → the functional group present (aldose/ketose) → ring structure formed Classification based on the number of carbon atoms No. of carbons Category name Examples 3 Triose Glyceraldehyde 4 Tetrose Erythrose 4  OUR LADY OF FATIMA UNIVERSITY COLLEGE OF PHARMACY PHARMACEUTICAL SEMINAR I 5 Pentose Ribose and Xylulose - Used to differentiate between blood types 6 Hexose Gluc, Fruc, Mann - Six membered cyclic form 7 Heptose Sedoheptulose - Aldohexose 8 Octose Glycero-manno- - Other names: Brain Sugar (Galactocerebroside) octulose - Rapidly absorbed in the small intestine 9 Nanose Neuraminic acid - C4 epimer of glucose Gal → Glu GALACTOSEMIA a condition which characterized by the galactose in the blood - Chmx test: Mucic acid test - (+) crystal formation  MANNOSE - Aldohexose - C2 epimer of glucose - Uses: Mannitol → Osmotic diuretic - Chxm test: Ozasone test - (+) needle-shaped crystal Hexoses Glucose Fructose Galactose Mannose Test for Hexoses Moore’s test Glucose Seliwanoff’s test Fructose BASED ON THE RING STRUCTURE FORMED Mucic acid test Galactose Monosaccharides containing 4 or more carbon atoms tend to have Osazone test Mannose cyclic structures Pentose Arabinose Gum arabic/Acacia Ribose RNA Deoxyribose DNA IMPORTANT INFO MALTOSE - sunflower shaped LACTOSE - powder puff shaped GALACTOSE - rhombic like  RIBOSE - Part of RNA, ATP, DNA - Five membered cyclic form - Aldopentose  GLUCOSE - Important sugar for genetics formation - Most abundant in nature - Nutritionally most important - Grape fruit good source of glucose (20-30% by mass) - Aldohexose or six membered cyclic form - Other names: Blood sugar (70-100 mg/100 mL), Physiologic sugar, Grape sugar, Dextrose, Corn sugar, D- glucopyanose  XYLOSE - Ketoaldose - Other names: Wood sugar  FRUCTOSE - Sweetest tasting of all sugars - Found in many fruits and in honey REDUCING PROPERTY OF MONOSACCHARIDES - Good dietary sugar due to higher sweetness - Five membered cyclic form Test Reagent Result - Ketohexose Fehling’s test Fehling’s A - Copper (+) Red - Other names: Levulose, Fruit Sugar sulfate - Source: Benedict’s test Copper citrate (+) Red Sucrose Tollen’s test Silver nitrate (+) Silver mirror Invert sugar Trommer’s test Alkaline solutions of (+) Red - Chmx test: Seliwanoiff’s test - (+) Red copper sulfate Nylander’s test Alkaline solution of (+) Black  GALACTOSE bismuth subnitrate - Synthesize in human 5  OUR LADY OF FATIMA UNIVERSITY COLLEGE OF PHARMACY PHARMACEUTICAL SEMINAR I Barfoed’s test Copper oxide (+) Red  GLYCOSIDE FORMATION REDUCING PROPERTY OF MONOSACCHARIDES  CYANOHYDRIN FORMATION - Also known as the ‘Kiliani Fischer’ synthesis - An organic chemical reaction by an aldehyde or ketone with a cyanide anion or a nitrile to form a cyanohydrin  AMINO ACID FORMATION  RUFF DEGRADATION - Removal of 1 carbon atom from molecule of an aldose - If one of the hydroxyl groups of a monosaccharide is - Ex. D-glucose to D-arabinose replaced with an amino group, an amino sugar is produced  LOBRY DE BRUYN DEGRADATION - Interconversion or transformation from aldose to ketose or from ketose to aldose, this process will then perform unsaturated alcohol  OSAZONE REACTION - The carbonyl group of Aldose will be linked to Phenylhydrazine - The product of the linkage is crystal called OSAZONE Mannosamine Galactosamine Glucosamine B.2. DISACCHARIDE  Contains 2 monosaccharide units-covalently bonded to each other  SUCROSE  OXIDATION REACTION - Other name: Table sugar - Oxidation means the addition of oxygen to a molecule or - Most abundant of all disaccharides and found in plants; it is the removal of hydrogen from a molecule produced commercially from the juice of sugar cane and - This will produce a sugar acid sugar beets  REDUICTION REACTION - Sugar cane contains up to 20% by mass sucrose - Reduction means the addition of hydrogen to a molecule or - Sugar beets contain up to 17% by mass sucrose the removal of oxygen from a molecule - Non-reducing sugar - Head to head linkages - GLU + FRU - C6H12O6 + C6H12O6 → C12H22O11 - Demulcent - Contained in Simple syrup NF → 85% w/v  MALTOSE - Other name: Malt sugar, Beer sugar - Digested easily by humans due to enzymes that can break alpha-1,4-linkaes but not beta-1,4-linkages of cellobiose. Therefore cellobiose cannot be digested by humans - Reducing sugar - GLU + GLU  CELLOBIOSE  PHOSPHORYLATION - Other name: Trehalose - The hydroxyl groups of monosaccharide can react with - Produced as an intermediate in the hydrolysis of the inorganic oxyacids to form inorganic esters polysaccharide cellulose - Contains two beta-D-glucose monosaccharide units linked through alpha-beta-1,4 glycosidic linkage - Non reducing sugar - GLU+ GLU  LACTOSE - Other name: Milk sugar - Made up of beta-D-galactose unit and a beta-D-glucose unit joined by a beta-1,4 glycosidic linkage 6  OUR LADY OF FATIMA UNIVERSITY COLLEGE OF PHARMACY PHARMACEUTICAL SEMINAR I - Reducing sugar  CELLULOSE - GLU+ GAL - β 1, 4 glycosidic bond - Lactulose - Linear homopolysaccharide with b (1→4) glycosidic bond → alkaline rearrangement of lactose - Up to 5000 glucose units with molecular mass of 900,000 → fructose and galactose amu → poorly absorbed - Cotton ~95% cellulose and wood ~50% cellulose → bacteria in the colon metabolize the disaccharide to - Humans don’t have enzymes that hydrolyze b (1→4) acetic and lactic acids so humans can not digest cellulose animals also lack → laxative effect these enzymes but they can digest cellulose because they have bacteria in their guts to hydrolyze cellulose B.3. OLIGOSACCHARIDE - It serves as dietary fiber in food readily absorbs water  Contains 2-10 monosaccharide units covalently bonded and results in softer stools - 20-35 of dietary fiber is desired everyday Sugar Sugar units Raffinose Gal + Glu + Fru  INULIN - diagnostic agent for kidney fxn Maltotriose Glu + Glu + Glu Sucralose Glu + Fru + Gal  CHITIN Gentianose Glu + Glu + Fru - Homopolyme of N-acetyl-B-D-glucosamine - Aminated glucose B.4. POLYSACCHARIDE - Similar to cellulose in both function and structure  Contains many monosaccharide units covalently bonded - Linear polymer with all b (1→4) glycosidic linkages it has a N-acetyl amino derivative of glucose Homoglycans Heteroglycans - Function is to give rigidity to the exoskeleton s of crabs, 1 component 2 or more component lobsters, shrimp, insects, and other arthropods Starch Gums Glycogen Mucilage  DEXTRAN Inulin - sucrose → dextrantransglucosylase enzyme system Dextran (dextran sucrase) Leuconostoc mesenteroides Cellulose - 10% solution = adjunct in the treatment of shock - Iron dextran inj. - hematinic  STARCH → AMYLOSE C. METABOLISM OF CARBOHYDRATES - linear - composed of 250 to 300 D-glucopyranose units PRIMARY PATHWAY FOR GLUCOSE MEABOLISM - linked by α -1, 4 glucosidic bonds - more soluble in water than amylopectin - amylose + iodine = deep blue complex → AMYLOPECTIN - branching (every 25 units) - consists of 1000 or more glucose units - α-1,4 linkages, α -1,6 (branch) - amylopectin + iodine = blue-violet or purple color IMPORTANT INFO Alpha -amylase (α -1,4-glucan 4-glucanohydrolase) → an enzyme present in pancreaticjuice and saliva, hydrolyzes starch by arandom splitting of α -1,4- glycosidiclinkages ß- Amylase (α- 1,4-glucan maltohydrolase) → produces its effect by removing maltose units from the nonreducing ends of polysaccharide molecules  METABOLISM → the end-product in the case of amylose is nearly pure - sum of all chemical reactions in order to maintain life maltose → Anabolism - building up  GLYCOGEN → Catabolism - α 1, 4 and α 1, 6 every 8-10 units - breaking down - Humans and animals storage polysaccharide - e.g. breakdown of proteins and amino acids, - Contains only glucose units glycogen to glucose, TAG to fatty acids - Branched chain polymer - α 1 → 4 glycosidic bonds in - 3 Stages of Catabolism straight chains and α 1 → 6 in branches 1. Hydrolysis of the complex molecules - Molecular mass: 3,000,000 (up to 1,000,000 glucose units) - the complex molecules are broken down - Three times more highly branched than amylopectin in into their component building blocks/simple starch molecule - Excess glucose in blood stored in the form of glycogen - e.g. proteins that are degraded into amino acids, polysaccharides to monosaccharides 2. Conversion of the building blocks to simple intermediates 7  OUR LADY OF FATIMA UNIVERSITY COLLEGE OF PHARMACY PHARMACEUTICAL SEMINAR I - the diverse building blocks that produced in IMPORTANT INFO stage 1 are further degraded to acetyl CoA, Catabolic Reactions also served to capture chemical few molecules, and some energy is captured energy in the form of ATP, particularly from the as ATP but the amount is small comparesd degradation of the energy-rich fuel molecules to energy produced in stage 3 3. Oxidation of Acetyl CoA - also referred to as tricarboxylic cycle or Krebs Cycle, which is the final common pathway in the oxidation of fuel molecules that is also produced by Acetyl CoA - will generate a large amount of ATP via oxidative phosphorylation, as electron flows from NADH to FADH2 → Amphibolic - combination of Catabolism and Anabolism - e.g. Kreb’s cycle IMPORTANT INFO Cellular Respiration - used convert food into energy using water and oxygen  ADENOSINE TRIPHOSPHATE  4 Major Pathways - “energy currency of the cell” because it traps energy 1. GLYCOLYSIS (happens in the cytosol) - links anabolic and catabolic reactions 2. INTERMEDIATE REACTION - composed of ADP + P + Energy Oxidation and Reduction 3. KREBS CYCLE (inside the mitochondrial membrane) 4. ELECTRON TRANSPORT CHAIN (inside the  OXIDATION AND REDUCTION mitochondrial membrane) Oxidation Reduction LEORA GEROA A. GLYCOLYSIS/EMBDEN-MEYERHOF PATHWAY (+) Oxygen (-) Oxygen  Happens in cytosol (-) Hydrogen (+) Hydrogen  Most common type of pathway and discovered by Gustav Catabolism Anabolism Embden and Otto Meyerhoff together with Jakub Karol Parnas → Embden Meyerhof Pathway allows the metabolic use of glucose to generate ATPs and then NADH as well as the  CO-ENZYMES several biosynthetic precursor-like pyruvates → NAD (Nicotinamide adenine dinucleotide) → Entener-Doudoroff Pathway -is the other pathway for - found in living organisms glycolysis - joined by PO4 groups  Glycolysis is important for the entry of glucose into the cells, - accepts 1 hydrogen (NADH) going to hepatic tissues via facilitated diffusion (having the - from Nicotinamide/Niacin/ Vit. B3 and Nicotinic acid passive transport of the molecules across the cell membranes → FAD (Flavin adenine dinucleotide) via specific membranes or proteins) - a redox cofactor that is related to many metabolic  Carrier mediated rxn  Mouth: Salivary Amylase (CHON) - acc3epts 2 hydrogens (FADH2)  Starch will not directly produce glucose - from Riboflavin/Vit. B2  Glycose + Lysis → Glucose breakdown → Co-enzyme A  Product of Digestion: Oligosaccharide (Dextrin)r mediated - precursor  End product: ATP and 2 Pyruvic acid - from Pantothenic acid/Vit. B5  Summary: - 1 Glu → 2 Pyruvic acid IMPORTANT INFO - 6 to 8 ATP Based on Electron transport chain  2 Kinds of Glycolysis:  1 NADH = 3 ATP 1. Aerobic (Mitochondria)  1 FAD = 2 ATP - involvement of oxygen; use to convert glucose to pyruvate and ATP, wherein pyruvates are burned for  FATES OF THE GLUCOSE energy, and then converted into fats via fatty acid → ATP production synthesis; could undergo TCA or KREBS CYCLE → Amino acid and triglycerides synthesis - the final product is pyruvate along with the production → Glycogen synthesis of eight ATP molecules → Digestion of Carbohydrates begins to digest the minute 2. Anaerobic (Cytoplasm) the food enters the mouth because the saliva secreted from - includes ATP production, and has recycled NADH by your salivary gland moistens the foods as it is chewed making the product lactate (muscles) → Saliva is important in the digestion of glucose because - the final product is lactate along with the production of saliva releases an enzyme called amylase, which begins two ATP molecules. the breakdown process of the sugars to carbohydrates → Ptyalin - is a starch hydrolyzing enzyme that is produced by the salivary glands, particularly from salivary amylase → Excess glucose is converted into glycogen which is sometimes stored in fats and proteins 8  OUR LADY OF FATIMA UNIVERSITY COLLEGE OF PHARMACY PHARMACEUTICAL SEMINAR I  Step 3: Phosphorylation using ATP - from fructose-6-phosphate to fructose-1,6- bisphosphate - 2nd mol of ATP is consumed (needed in the addition of PO4 group to fructose-6-phosphate) - Enzyme: PHOSPHOFRUCTOKINASE (PFK) ; rate-limiting enzyme; requires Mg2+ ion for its activity - at this point, Glucose-6-phosphate & Fructose-6- phosphate can enter other metabolic pathways, but Fructose-1,6-biphosphate can only enter glycolysis - Product: FRUCTOSE-1,6-BISPHOSPHATE - reaction is irreversible. - one ATP is utilized for phosphorylation. - second priming reaction IMPORTANT INFO  Glycolysis is composed of 10 processes and divided into 2 Steps 4-10 are considered as “Energy Generating Stage”, stages wherein ATP is used in the process; where the conversion 1. Preparatory Phase (Steps 1-5) of pyruvate happens → STEP 1-3: Energy Consuming Stage - it uses ATP instead of producing ATP; aka “PREPARATORY PHASE”  Step 4: Formation of Cleavage (Formation of 2  Step 1: Phosphorylation using ATP Triose Phosphate) - glucose ring is phosphorylated (added PO4 - Enzyme: ALDOLASE is responsible for group to a molecule that is derived from ATP) catalyzing the cleavage of Fructose-1,6- biPO4 - 1 mol ATP consumed (conversion of ATP to (FBP) to yield two 3-carbon molecules ADP) - the structure of Fructose-1,6-biphosphate will be - Enzyme: HEXOKINASE is responsible for divided into two: phosphorylation and requires Magnesium for its  1 mol contains a ketone group → activity (enzyme that ends with -KINASE means an Dihydroxyacetone PO4 (DHAP) enzyme that phosphorylates other molecules)  other mol contains an aldehyde group → - Product: GLUCOSE-6-PHOSPHATE (at 6 carbon GAP attachment of PO4 group) - Products: GLYCERALDEHYDE-3-PHOSPHATE - First priming reaction (GAP) **this molecule can already proceed to the glycolytic pathway; DIHYDROXYACETONE PHOSPHATE (DHAP) **this molecule is further acted by triphosphate isomerase (TIM) to produce or reorganize DHAP to GAP, due to the ketone group present in DHAP - reaction is reversible. - this reaction produces two 3-carbon molecules but not yet fully converted into pyruvate  Step 2: Isomerization - Formation of fructose-6-phosphate - Enzyme: PHOSPHOGLUCOSE ISOMERASE (PI) or PHOSPHOHEXOSE ISOMERASE - Rearrangement of the carbon-oxygen bond (from 6-membered ring to 5-membered ring) from Aldose to Ketose - Carbon 1 of Glucose is no longer part of the ring structure Product: FRUCTOSE-6-PHOSPHATE 2. Payoff Phase  Step 5: Isomerization (Formation of Glyceraldehyde-3-PO4) - conversion of DHAP into GAP - this is to further proceed into the glycolysis process - Ketone → Aldehyde - Enzyme: TRIPHOSPHATE ISOMERASE aka triose-phosphate isomerase 9  OUR LADY OF FATIMA UNIVERSITY COLLEGE OF PHARMACY PHARMACEUTICAL SEMINAR I - GAP is on the direct pathway of glycolysis,  Step 7: Phosphorylation of Adenosine whereas DHAP is not. Hence, Triphosphate Diphosphate (Formation of 3- isomerase converts DHAP into GAP, which is Phosphoglycerate) useful for generating ATP. Thus, the net result is - 1,3-biphosphoglycerate is converted to 3- that glucose is now cleaved into 2 molecules of PHOSPHOGLYCERATE glyceraldehyde-3-phosphate - ATP-producing step - reaction is rapid and reversible - ENZYME: PHOSPHOGLYCERO KINASE - this stage is responsible for the reason why the - there is a loss of phosphate group from the end-product of glycolysis has 2 pyruvates starting material (1,3-biphosphoglycerate) - Product: another GLYCERALDEHYDE-3- - the loss phosphate is transferred to a molecule PHOSPHATE (GAP) of ADP that yields the first ATP in this process - at this point, 2 molecules of GAP were already - since we have two molecules of 1,3- produced to enter step 6 biphosphoglycerate, the product has two ATPs. - From this point forward, there would Although these ATPs produced will be canceled be 2 products yielded in every out to recompense the ATPs that were used in step (STEPS 5-10) steps 1 and 3. Therefore, the net ATP equates to zero (0) - the enzyme transfers the high-energy phosphoryl group from the carboxyl group of 1,3- bisphosphoglycerate to ADP, forming ATP and 3- phosphoglycerate - this is a unique example where ATP can be produced at the substrate level without participating in the electron transport chain. This type of reaction where ATP is formed at substrate level is called Substrate level phosphorylation  Step 6: Oxidation and Phosphorylation using Inorganic Phosphate - formation of 1,3-biphosphate glycerate - contains TWO MAIN EVENTS:  GAP is oxidized by the coenzyme Nicotinamide Adenine Dinucleotide (NAD)  The molecule is phosphorylated by the  Step 8: Isomerization (Formation of addition of a free phosphate group Phosphoglycerate) - ENZYME: GLYCERALDEHYDE-3-PHOSPHATE - rearrangement of the phosphate group on 3- DEHYDROGENASE (GAPDH) that allows phosphoglycerate molecules NAD to pull the Hydrogen atom off GAP → NADH, - the phosphate group positioned on the 3rd producing 3 ATPs. The phosphate groups carbon will be shifted to the 2nd carbon attack the GAP molecule and release it from the - PRODUCT: 2-PHOSPHOGLYCERATE enzyme to yield the final product. - ENZYME: PHOSPHOGLYCERATE MUTASE - Product: 1,3-BIPHOSPHOGLYCERATE, NADH, (PGM) or PHOSPHOGLYCEROMUTASE Hydrogen atom (GAP + NAD + Pi ⇌ 1,3- - Mutase catalyzes the transfer biphosphate + NADH + H+) of a functional group from one position - 1,3-BIPHOSPHOGLYCERATE is an to another; a readily reversible example of a high-energy phosphate reaction; Mg2+ is essential for this group. This is the energy that is used to reaction convert ADP to ATP - an energy-yielding reaction. Reactions of this type in which an aldehyde group is oxidized to acid are accompanied by the liberation of large amounts of potentially useful energy. During this reaction, NAD+ is reduced to NADH - reversible reaction  Step 9: Dehydration (Formation of Phosphoenolpyruvate) - conversion of 2-phosphoglycerate to phosphoenolpyruvate (PEP) - ENZYME: ENOLASE or PHOSPHOPYRUVATE HYDRATASE - the second step where high-energy-rich compounds are produced, with step 6 being the first one - PRODUCT: PHOSPHOENOL PYRUVATE (PEP) - high-energy-rich compound aside from step 6 10  OUR LADY OF FATIMA UNIVERSITY COLLEGE OF PHARMACY PHARMACEUTICAL SEMINAR I - the reaction is reversible - NADH produced in the cytosol, therefore, it cannot DIRECTLY enter the mitochondrial matrix but the 2 electrons of NADH can enter the mitochondrial matrix from the cytosol by using the substrate shuttles - NADH cannot easily enter the mitochondrial matrix because: 1. NADH is produced from  Step 10: Phosphorylation of ADP (Formation CYTOSOL of Pyruvate) 2. the mitochondria have the - ATP-producing step barrier that separates the - last step in glycolysis inner membrane to the - conversion of phosphoenolpyruvate into intermembrane space (the pyruvates outside portion of the - ENZYME: PYRUVATE KINASE mitochondria) - there is a transfer of phosphate group; the - Examples of Substrate Shuttle: phosphate group attached to the 2nd carbon of 1. Glycerol PO4 shuttle phosphoenolpyruvate is transferred to the - brain & skeletal muscles molecule of ADP to yield ATP - 2 electrons are transferred - step that produced 2 molecules of ATPs due to 2 from NADH to molecules of pyruvate dihydroxyacetone PO4 (DHAP) - PRODUCT: PYRUVATE with the help of the enzyme - first PEP is made into a transient intermediary glycerophosphate of enol pyruvate; which is spontaneously dehydrogenase, resulting a isomerized into keto pyruvate, the stable form of synthesis of 2 ATPs for every pyruvate cytosolic NADH synthesized - the pyruvate kinase is a key glycolytic enzyme. 2. Malate-Aspartate shuttle This step is irreversible - heart, liver, & kidneys - produces NADH and yields 3 ATPs on each cytosolic matrix oxidized by malate dehydogenase Summary of ATPs → 2 ATPs were consumed → 4 ATPS were generated (net gained) → 1 glucose = 2 Pyruvate and 2 ATPs Step Reactions and ATPs 1 Glucose → Glucose-6-phosphate ATP: -1 3 Fructose-6-phosphate → Fructose-1,6-bisphosphate ATP: -1 7 2 mol of 1,3-biphosphoglycerate → 2 mol of 3- phosphoglycerate Substrate-level Phosphorylation Oxidative Phosphorylation ATP: +2 - No middle man (happens when - Have middlemen in the form of 10 2 mol of phosphoenolpyruvate → 2 mol of pyruvate ADP is converted to ATP by the NADH & ETC to produce the ATP ATP: +2 direct transfer of PO4 group 2) - The phosphate comes from a Total ATPs = +2 - Direct process → Direct pool of inorganic PO4 instead of trasnfer – the phosphate group is directly from another molecule WHAT WILL HAPPEN TO NADH? donated & transferred from a (The energy needed to phosphorylated intermediate (e.g. phosphorylate ADP comes from a  OXIDATIVE PHOSPHORYLATION step 7) proton gradient together with ETC → a biochemical process by which ATP is synthesized from - LOCATION: Cytoplasm & NADH) ADP, a result of the transfer of electrons & H+ ions - LOCATION: Inner mitochondrial from NADH or FADH2 to Oxygen through to the electron membrane carrier involved in that electron transport chain producing ATP B. INTERMEDIATE REACTION: FORMATION OF ACETYL CoA → metabolic pathway wherein the cell used these enzymes to oxidize the nutrients, thereby, releasing energy which is used to reform ATP → IN THE EUKARYOTES: - Oxidative phosphorylation happens inside the mitochondria (because the inner mitochondrial membrane lack NADH transporter & the NADH is produced in the cytosol) 11  OUR LADY OF FATIMA UNIVERSITY COLLEGE OF PHARMACY PHARMACEUTICAL SEMINAR I  occurs in Mitochondria in close proximity to ETC to oxidize the reduced co-enzymes NADH and FADH2  series of chemical reactions that are used by all aerobic organisms used to generate energy through the oxidation of acetate (the product Acetyl CoA was from the 2 pyruvates produced after the glycolysis) that is derived from CHO, fats & CHON to be converted to CO2 & chemical energy in the form of ATP  PURPOSE: Burn the Acetyl CoA that is made from fats, glucose, & proteins to be able to make the ATPs incorporation with the oxidative phosphorylation  GOAL: To produce CO2  Composed of 8 steps and happens in Mitochondria  Step 1: Formation of Citrate - process involved is CONDENSATION  Aerobic reaction - Acetyl CoA will undergo Krebs Cycle by the PROCESS OF → undergo oxidative decarboxylation of pyruvate CONDENSATION and will donate acetyl group to the 4 → ENZYME: Pyruvate dehydrogenase complex carbon OXALOACETATE by the CITRATE SYNTHASE to → PRODUCT: 2 mol of Acetyl CoA that would enter form a PRODUCT: CITRATE (6 achiral carbon) Citric acid cycle or Krebs Cycle (oxidative decarboxylation)  ACETYL CoA → Acetyl CoA product is ready for Kreb’s cycle & Fatty Acid - major fuel for Kreb’s cycle synthesis - derived from oxidative decarboxylation of → PRODUCES 2 Acetyl CoA = 2 NADH = 2x3 = 6 ATPs pyruvate  Anaerobic reaction → WITHOUT the presence of OXYGEN  Step 2: Formation of Isocitrate → conversion of pyruvic acid/pyruvate to lactic acid - via the process of ISOMERIZATION → ENZYME: lactate dehydrogenase - Citrate is isomerized to ISOCITRATE → PRODUCT: lactic acid - 1st step to produce NADH and CO2 - when lactic acid is produced in the body during - Citrate with the aid of the ACONITASE by the process of intense exercise the lactate accumulates in the dehydration produces CIS-ACONITASE and with the muscles causing a drop in the intracellular pH & aid of the ACONITASE by the process of hydration potentially result in MUSCLE CRAMPS. Hence, ISOCITRATE (chiral cmpd) present in muscle fatigue - if diffuses in the bloodstream it can be used by  Step 3: Oxidation of Isocitrate and Formation of CO2 the liver to make glucose - via OXIDATION AND DECARBOXYLATION - if there is a level of lactate/elevated - Isocitrate is oxidized & decarboxylated by ISOCITRATE concentration especially in the plasma is called DEHYDROGENASE to alpha-ketoglutarate producing LACTIC ACIDOSIS NADH & CO2 - Isocitrate acted upon by the isocitrate IMPORTANT INFO dehydrogenase [NAD is converted to its reduced Acetyl CoA will now enter the mitochondria for Kreb’s form NADH (NAD to NADH + H+)] → forming cycle OXALOSUCCINATE producing → H+ CO2 and alpha-ketoglutarate C. KREB’S CYCLE  Step 4: Oxidation of Alpha-ketoglutarate and Formation of CO2 - via OXIDATION AND DECARBOXYLATION - 2nd step to produce NADH and CO2 - alpha-ketoglutarate is oxidatively decarboxylated to SUCCINYLY COA by ALPHA-KETOGLUTARATE DEHYDROGENASE COMPLEX  Alpha-ketoglutarate - an enzyme very similar to pyruvate dehydrogenase - activated by CALCIUM and inhibited by NADH & succinyl CoA - alpha-ketoglutarate + NAD + H+ + CoA-SH aided by the alpha-ketoglutarate dehydrogenase complex producing → succinyl CoA + NADH + CO2 + H+  Step 5: Thioester bond cleavage in Succinyl CoA and Phosphorylation of GDP - via PHOSPHORYLATION  “Tricarboxylic acid cycle” or “ Citric acid cycle” named after Hans - a substrate-level phosphorylation Adolf Krebs - step-producing GTP  an aerobic pathway because it requires oxygen - Succinyl CoA is cleaved by SUCCINYL THIOKINASE OR  final pathway where the oxidative catabolism of carbohydrates, SUCCINYL COA SYNTHETASE producing SUCCINATE amino acids, and fatty acids converge their carbon skeleton AND GTP (Guanosine triphosphate) being converted to CO2 12  OUR LADY OF FATIMA UNIVERSITY COLLEGE OF PHARMACY PHARMACEUTICAL SEMINAR I - Succinyl CoA acted by the enzyme succinyl CoA  every oxidation of something must be coupled to the reduction of synthetase forming succinyl phosphate then acted something else. Then, the molecule or atom that loses an again by the enzyme succinyl CoA synthetase electron has been oxidized & the one that gains them has been producing GTP and SUCCINATE reduced (Vi LeORA & Vd GeROA) - Succinyl CoA releases energy, which is trapped → Oxidants/Oxidizing agents by the formation of GTP - molecules or compounds that oxidizes other - The fxn of GTP is similar to ATP which is to compounds or molecules; undergone reduction or being STORE energy in the form of a high energy reduced in the process phosphate bond - examples:  Lactate oxidized to pyruvate IMPORTANT INFO  NADH (reduced form) oxidized to NAD Step 6-8 is a sequence of functional group changes → Reductants/Reducing agents - compounds that reduced other compounds;  Step 6: Oxidation of Succinate substances that undergone oxidation process - via OXIDATION - examples: - step-producing FADH2  Pyruvate reduced to lactate - succinate is oxidized to FUMARATE by SUCCINATE  NAD reduced to NADH DEHYDROGENASE producing FADH2 - Succinate aided by succinate dehydrogenase to IMPORTANT INFO produce FUMARATE + FADH2 Pyruvate and NAD - oxidizing agent ** FAD is reduced to FADH2 NADH and Lactate - reducing agent  Step 7: Hydration of Fumarate  as the fuel molecules are oxidized the electrons that have been - via HYDRATION lose are used to make NADH & FADH2 - Fumarate is hydrated to MALATE by FUMARASE  the fxn ETC & Oxidative Phosphorylation is to take electrons from molecules (NADH & FADH2) & transfer them to oxygen  Step 8: Oxidation of L-Malate to Regenerate making ATP in the process Oxaloacetate  Location: inner mitochondrial membrane - via OXIDATION  Flavin adenine dinucleotide or FADH2 - 3rd step to produce NADH - FADH2 is only produced in Kreb’s cycle - Malate is oxidized to OXALOACETATE by MALATE  Nicotinamide adenine dinucleotide (NADH) DEHYDROGENASE producing NADH - NADH is a product of both the glycolysis and Kreb cycles  O2 - final electron acceptor Summary of ATPs  Complete O2 reduction → H2O Step 3,4,8 3 NADH 9 ATP  Partial O2 reduction → superoxide anioons (O2), hydroxyl Step 6 1 FADH2 2 ATP radicals (OH-) and peroxides (H2O2) Step 5 1 GTP 1 ATP  Products: ATP + H2O 12 ATPs per cycle x  Mobile electron carriers: Cytochrome Q and C 2 due to two  4 Protein Complexes are responsible for ETC by transporting pyruvates that will electrons to acquire energy enter the cycle.  Complex I: NADH Coenzyme Q reductase TOTAL: 24 ATPs - for every 2 electron = 4 protons are pump - inhibited by rotenone which is a rat poison IMPORTANT INFO - largest protein complex containing 40 sub-units including Flavin mononucleotide (FMN ) or Iron sulfur GDP/GTP binding protein activity is regulated by the binding & hydrolysis of GTP. These proteins are crucial for protein (FeSP) signal transduction events resulting in cell division, - received electrons coming from NADH (from sugar cytoskeleton management, and sensory perception. metabolism) that will go to ETC to deposit 2 high energy electrons to COMPLEX I; when these small electrons pass through complex they will pass Summary of the Pathways through a long chain of redox centers (sub-units) and Pathway No. of ATP produced when they finally exit complex I it will donate the 2 Glycolysis (Substrate-level  Substrate-level electrons to a coenzyme Q molecule that will transfer Phosphorylation and Oxidative Phosphorylation = 2 ATPs it to complex III Phosphorylation)  Oxidative Phosphorylation  COENZYME Q comes from a QUINONE = 4-6 ATPs molecule containing 10 isoprene units TOTAL: 6-8 ATPs attached to the quinone units Intermediate Pathway  2 Acetyl CoA = 2 NADH = 2 - STEP 1: Interaction of NADH with FMN will oxidize X 3 = 6 ATPs the NADH to NAD and passes 2 H+ & 2 electrons to TOTAL: 6 ATPs the FMN which is also reduced to FMN2 Kreb’s cycle  24 ATPs - last step: re-conversion of FeSP TOTAL: 36-38 ATPs  Complex II: Succinate–coenzyme Q reductase D. ELECTRON TRANSPORT CHAIN - PURPOSE: process the FADH2 from the citric acid  electrons usually are not floating around the space particularly in cycle/Krebs cycle the Mitochondria they are stuck in some atoms or others - smaller consisting 4 sub-units including 2 FeSP [consequence - when one thing losses ELECTRONS something - high-energy FADH2 molecules will enter carrying else must gain them] (OXIDATION & REDUCTION is a electrons to pass through the redox centers or sub- simultaneous process; if one molecule is oxidized other will be units inside complex II and when it finally exit reduced in simultaneous procedure or vice versa) 13  OUR LADY OF FATIMA UNIVERSITY COLLEGE OF PHARMACY PHARMACEUTICAL SEMINAR I complex II, it will donate them to coenzyme Q that  During electron transfer at the three classic sites of will transfer the electrons to complex III phosphorylation particularly Complex I, II, and III → the protons are pumped out the Mitochondria to the Cytoplasm. Thus, proton  Complex III: Coenzyme Q–cytochrome C reductase pumping makes the mitochondria ALKALINE - for every 2 electron = 4 protons are pump  F1FO ATPase is an enzyme that allows the protons back to the - inhibited by Antimycin A Mitochondria (from the intermembrane space back to the inner - has 11 different sub-units & also includes FeSP and mitochondrial matrix) cytochromes  Inhibitors can block the flow of electrons at the specific site that  Cytochrome is a heme-containing protein in can also inhibit the electron flow and ATP synthesis which reversible oxidation & reduction of an iron atom occur Different Inhibitors - the electron will enter complex III and will pass through Inhibitor Site Effect redox centers before reaching cytochrome C that will Cyanide Cytochrome oxidase Blocks transfer of electrons deliver electrons from complex III to complex IV to O2. Blocks at site III. Antimycin Electron transfer from All intermediates before and  Complex IV: Cytochrome C oxidase cyt b to cyt c1 including cyt a will be in the - for every 2 electron = 2 protons are pump reduced state; all - inhibited by cyanide & CO intermediates after and - has 13 sub-units including 2 cytochromes including cyt c1 will be in the - end of ETC oxidized state. Blocks at site - electrons enter, where the 4 electron converts a II. molecule to Oxygen to produce 2 molecules of water Retenone NADH-CoQ Blocks oxidation of NADH reductase (site 1). NADH will become IMPORTANT INFO reduced; substrates such as The presence of Oxygen is important in the ETC succinate that enter via especially in complex IV because oxygen will serve as the FADH will still be oxidized final acceptor of electrons in the last complex. Therefore, and make 2 ATPs/mol. absence of Oxygen in ETC comes to a halt & the ATP Oligomycin ADP phosphorylation Block phosphorylation of synthesis will also stop ADP. Does not inhibit uncoupled oxidations. All the complexes are proton pums except for Complex II Atracrtyloside ADP-ATP transporter Inhibits entry of ADP into and mitochondria and ATP bongkrekate export. Stops electron transport because of lack of ADP. Inside, all ADP is converted to ATP. SECONDARY PATHWAY FOR GLUCOSE METABOLISM A. HEXOSE MONOPHOSPHATE SHUNT (HMP SHUNT)/PPP  “Phosphogluconate Pathway”  NADH is required for the biosynthesis of fatty acid (particularly NADPH)  PURPOSE: It makes NADPH for biosynthesis of fatty acids and riboses for DNA and RNA synthesis  NADPH is a reducing agent that is reserved for biosynthetic pathway particularly in the FA synthesis. Thus, HMP shunt is called upon when reducing equivalents and fatty acids synthesis is turned on  NADPH is also used to keep the cellular and mitochondria glutathione in the reduced form through the action of the glutathione reductase  Ribose-5-Phosphate  D-ribose is used for the synthesis of nucelic acid  Occurs in the cytosol  The inner mitochondrial matrix consists of protons that will be - to produce ribose-5-PO4 for nucelotide/DNA synthesis transported or pump from the inner mitochondrial membrane out - to produce NADPH from NADP+ for FA and steroid synthesis, to the intermembrane space maintaining reduced Gluthathione inside RBC  Protein complexes are used to produced protons from inner - to interconvert pentose and hexoses mitochondrial membrane to intermembrane space, particularly  Enzyme: Glucose-6-PO4 dehydrogenase