16.1 Intro to Proteins Proteins are polymers in which the monomer units are amino acids. The name “protein” comes from the Greek, and means “of first importance.” Proteins are the most abundant biomolecules in animals (including humans) and have the widest variety of structures. Proteins contain nitrogen; carbohydrates and lipids generally do not. Copyright © Houghton Mifflin Company. All rights reserved. 3–1 16.2 Amino Acids An amino acid contains both an amino group (–NH2) and a carboxylic acid (–COOH). Both groups are on the same carbon, the -carbon. The carbon also carries an R group and a hydrogen atom. O H N H C H Copyright © Houghton Mifflin Company. All rights reserved. C H O R 3–2 16.2 Amino Acids There are 20 standard amino acids, each with a different R group. There are four categories of amino acids. Copyright © Houghton Mifflin Company. All rights reserved. 3–3 16.2 Amino Acids Polar neutral amino acids. Copyright © Houghton Mifflin Company. All rights reserved. 3–4 16.2 Amino Acids Polar acidic and basic amino acids. Copyright © Houghton Mifflin Company. All rights reserved. 3–5 16.2 Amino Acids Polar acidic and basic amino acids. Copyright © Houghton Mifflin Company. All rights reserved. 3–6 16.2 Amino Acids Ten amino acids cannot be produced by the body, and must be obtained through the diet. These are the essential amino acids. Copyright © Houghton Mifflin Company. All rights reserved. 3–7 16.3 Handedness The -carbon in an amino acid is chiral. Most amino acids have the L-configuration. Copyright © Houghton Mifflin Company. All rights reserved. 3–8 16.4 Acid-Base Properties Amino acids undergo intramolecular proton transfer. They always exist as zwitterions (double, or hybrid) ions. O H N H C C H H H O R Neutral Molecule O H N C C O H H R Zwitterion Zwitterions have no net electrical charge. Copyright © Houghton Mifflin Company. All rights reserved. 3–9 16.4 Acid-Base Properties In acidic solution, the carboxylate group is protonated. This produces a cation. H O H N C + H3O C O H H R Zwitterion + acid Copyright © Houghton Mifflin Company. All rights reserved. H O H N C + H2O C OH H H R Positive net charge 3–10 16.4 Acid-Base Properties In basic solution, the ammonium group is deprotonated. This produces an anion. H O H N C + OH C O H H R Zwitterion + base Copyright © Houghton Mifflin Company. All rights reserved. O H N C + H2O C O H H R Negative net charge 3–11 16.4 Acid-Base Properties The pH at which an amino acid exists as its uncharged zwitterion is called the isoelectric point. Neutral amino acids have isoelectric points about pH 6. Acidic amino acids have low isoelectric points, because the carboxylate group must be protonated. Basic amino acids have high isoelectric points, because the amino group must be deprotonated. Copyright © Houghton Mifflin Company. All rights reserved. 3–12 16.5 Peptide Formation Amino acids condense to form amide, or peptide, bonds. The reactions are catalyzed by enzymes. H H H N C H R1 H O + C H O H H H N C H R1 a dipeptide Copyright © Houghton Mifflin Company. All rights reserved. H O N C H R2 enzyme C O O H C N C H R2 O C + H2O O 3–13 16.5 Peptide Formation A chain of any length can form. A polypeptide contains 50 or fewer amino acid residues. A protein contains more than 50 amino acid residues. Central Dogma of Molecular Biology Protein synthesis is directed by RNA (ribonunucleic acid). DNA (deoxyribonucleic acid) acts as a template for RNA synthesis. Copyright © Houghton Mifflin Company. All rights reserved. 3–14 16.6 - 16.10 Protein Structure The structure of proteins and peptides is critical to their function in organisms. It is divided into four levels. Primary structure of proteins refers to the sequence of amino acid residues. Copyright © Houghton Mifflin Company. All rights reserved. 3–15 16.6 - 16.10 Protein Structure The structure of proteins and peptides is critical to their function in organisms. It is divided into four levels. Primary structure of proteins refers to the sequence of amino acid residues. The illustration shows human myobglobin. Copyright © Houghton Mifflin Company. All rights reserved. 3–16 16.6 - 16.10 Protein Structure Secondary structure of proteins refers to the arrangement adopted by the backbone portion of the protein. It results from hydrogen bonding between N–H and C=O groups. Two major secondary structures are seen: the -helix, formed by a coiled chain the -pleated sheet, formed by hydrogen bonds between extended chains or segments of a single chain Copyright © Houghton Mifflin Company. All rights reserved. 3–17 16.6 - 16.10 Protein Structure Views of the -helix, formed by a coiled chain Copyright © Houghton Mifflin Company. All rights reserved. 3–18 16.6 - 16.10 Protein Structure Views of the -pleated sheet, formed by hydro-gen bonds between extended chains or seg-ments of a single chain Copyright © Houghton Mifflin Company. All rights reserved. 3–19 16.6 - 16.10 Protein Structure Views of the -pleated sheet, formed by hydro-gen bonds between extended chains or seg-ments of a single chain Copyright © Houghton Mifflin Company. All rights reserved. 3–20 16.6 - 16.10 Protein Structure Tertiary structure refers to the overall three-dimensional shape of a protein. This illustration shows the structure of myoglobin. Copyright © Houghton Mifflin Company. All rights reserved. 3–21 16.6 - 16.10 Protein Structure Four types of attractive forces give rise to the tertiary structure of proteins. Disulfide bonds Electrostatic interactions, a.k.a. salt bridges Hydrogen bonds Hydrophobic interactions Copyright © Houghton Mifflin Company. All rights reserved. 3–22 16.6 - 16.10 Protein Structure Disulfide bonds form between –SH groups of cysteine residues. They are covalent bonds. C O H C CH2 SH + HS H N Copyright © Houghton Mifflin Company. All rights reserved. O C CH2 C H N H C O H C CH2 S H N enzyme O S C CH2 C H N H 3–23 16.6 - 16.10 Protein Structure Disulfide bonds between the two chains of human insulin. Copyright © Houghton Mifflin Company. All rights reserved. 3–24 16.6 - 16.10 Protein Structure Electrostatic interactions between side-chain carboxylate and ammonium ions are ionic bonds. They are also called salt bridges. Copyright © Houghton Mifflin Company. All rights reserved. 3–25 16.6 - 16.10 Protein Structure Hydrogen bonds form between hydroxyl and amide functional groups on side chains of amino acid residues. Copyright © Houghton Mifflin Company. All rights reserved. 3–26 16.6 - 16.10 Protein Structure Hydrophobic interactions occur between nonpolar side chains on amino acid residues. They involve dispersion forces similar to those that form micelles. Copyright © Houghton Mifflin Company. All rights reserved. 3–27 16.6 - 16.10 Protein Structure Quaternary structure refers to the arrangement of polypeptide chains within a protein that are not covalently bound to each other. The chains are bound by the same forces that give rise to tertiary structure. The next slide shows the quaternary structure of hemoglobin. Copyright © Houghton Mifflin Company. All rights reserved. 3–28 16.6 - 16.10 Protein Structure Quaternary structure refers to the arrangement of polypeptide chains within a protein that are not covalently bound to each other. Copyright © Houghton Mifflin Company. All rights reserved. 3–29 16.11 Protein Classification Proteins can be classified by composition or by morphology (shape/structure). Classifications based on composition: Simple proteins contain only amino acid residues. More than one chain may be present. Conjugated proteins have prosthetic groups, components that are not made up of amino acids. These can be organic or inorganic. Copyright © Houghton Mifflin Company. All rights reserved. 3–30 16.11 Protein Classification Types of conjugated proteins: Copyright © Houghton Mifflin Company. All rights reserved. 3–31 16.11 Protein Classification Classifications based on morphology: Fibrous proteins are rod-shaped or stringlike. They have structural or movement functions. They have very long chains. They are not water-soluble. Globular proteins are “globby.” They have functions other than structure or movement. They have chains of moderate length. They dissolve in water or form colloids. Copyright © Houghton Mifflin Company. All rights reserved. 3–32 16.11 Protein Classification Types of fibrous and globular proteins: Copyright © Houghton Mifflin Company. All rights reserved. 3–33 16.11 Protein Functions 1. Catalysis: Proteins called enzymes catalyze biochemical reactions. 2. Structure: Proteins are the main structural molecules in animals. Collagen is found in skin, bone, connective tissue (tendons, etc.) Keratin in hair, nails, skin Copyright © Houghton Mifflin Company. All rights reserved. 3–34 16.11 Protein Functions 3. Storage: Proteins provide a way to store small molecules or ions in the organism. Ovalbumin stores amino acids in bird eggs. Casein stores amino acids in milk. Ferritin stores iron ions in animal spleens. Copyright © Houghton Mifflin Company. All rights reserved. 3–35 16.11 Protein Functions 4. Protection: Antibodies form complexes with foreign protein from viruses and bacteria, and help destroy them. Fibrinogen and thrombin are proteins involved in blood clot formation. Copyright © Houghton Mifflin Company. All rights reserved. 3–36 16.11 Protein Functions 5. Regulation: Hormones trigger a specific process in target tissues. Many are proteins or peptides. Insulin regulates glucose metabolism. Vasopressin regulates volume of urine and blood pressure. Oxytocin regulates contraction of the uterus and lactating mammary glands. Copyright © Houghton Mifflin Company. All rights reserved. 3–37 16.11 Protein Functions 6. Nerve Impulse Transmission: Some proteins act as receptors of small molecules that pass across synapses, gaps between nerve cells. Rhodopsin is a protein in the retina. It is activated by isomerization of retinal, a molecule derived from Vitamin A. Copyright © Houghton Mifflin Company. All rights reserved. 3–38 16.11 Protein Functions 7. Movement: Proteins in muscle are responsible for contraction and relaxation. Actin and myosin are fibrous proteins that slide across each other in muscle movement. Copyright © Houghton Mifflin Company. All rights reserved. 3–39 16.11 Protein Functions 8. Transport: Proteins transport ions and molecules through the blood stream and other body fluids. Hemoglobin transports oxygen through the blood stream. Serum albumin transports fatty acids through the blood stream. Lipoproteins transport lipids through various body fluids. Copyright © Houghton Mifflin Company. All rights reserved. 3–40 16.12 Protein Deactivation Proteins can be deactivated by two processes, hydrolysis and denaturation. Hydrolysis is the reverse of peptide bond formation. It reduces a protein to smaller polypeptide molecules and free amino acids. Hydrolysis can be caused by enzymes or strong acids or bases. It is part of the normal digestion of proteins in the stomach. Copyright © Houghton Mifflin Company. All rights reserved. 3–41 16.12 Protein Deactivation Hydrolysis of a tripeptide to amino acids: H H H H N C H R1 H H N C H R1 O H C N C H R2 O O + C H C H O Copyright © Houghton Mifflin Company. All rights reserved. N C H R3 H H N C H R2 H2O acid, base, or enzyme O C O O + C O H H H N C H R3 O C O 3–42 16.12 Protein Deactivation Denaturation is the partial or complete disorganization of a protein’s threedimensional shape. Denaturation is caused by disruption of the attractive forces that produce this shape. Heat, acids or bases, detergents, organic solvents, ions of heavy metals, and violent whipping or shaking can denature proteins. Copyright © Houghton Mifflin Company. All rights reserved. 3–43 16.12 Protein Deactivation Denaturation of a protein: Copyright © Houghton Mifflin Company. All rights reserved. 3–44 16.13 Enzymes Enzymes are catalysts for biochemical reactions. Most are globular proteins, although a few are ribonucleic acids. Catalysts increase the rate of a chemical reaction, but they are not consumed in the reaction. Enzymes can increase the rate of a reaction by a factor of 10 20 compared to that of the uncatalyzed reaction. Copyright © Houghton Mifflin Company. All rights reserved. 3–45 16.13 Introduction to Enzymes Enzyme catalysis has three key characteristics: Efficiency Reactions are very fast under mild conditions. Specificity Enzymes catalyze reactions of just one compound, or a few similar compounds (substrates). Regulation Catalysis can be controlled by the cells in which reactions occur. Copyright © Houghton Mifflin Company. All rights reserved. 3–46 16.14 Names and Classification The name of a specific enzyme has three parts and ends in “-ase.” H2N O C Urea amidohydrolase NH2 1 2 1. Substrate 2. Functional Group 3. Reaction Copyright © Houghton Mifflin Company. All rights reserved. 3 2 NH3 + CO2 Urea Amide Hydrolysis 3–47 16.14 Names and Classification Enzymes are named by the type of reaction they catalyze: 1. Oxoreductases Oxidation-reduction reactions 2. Transferases Move functional groups 3. Hydrolases Hydrolysis reactions 4. Lyases Addition across double bonds or the reverse (elimination) 5. Isomerases Rearrange the substrate 6. Ligases Formation of bonds with ATP cleavage Copyright © Houghton Mifflin Company. All rights reserved. 3–48 16.14 Names and Classification Like other proteins, enzymes can be simple or conjugated. Simple enzymes contain only amino acid residues. More than one chain may be present. Conjugated enzymes have components that are not made up of amino acids. These can be organic or inorganic. Copyright © Houghton Mifflin Company. All rights reserved. 3–49 16.14 Names and Classification Conjugated enzymes consist of two parts: An apoenzyme is the protein portion of a conjugated enzyme. A cofactor is the nonprotein portion of a conjugated enzyme. There are three types: Prosthetic groups Coenzymes Minerals Copyright © Houghton Mifflin Company. All rights reserved. 3–50 16.14 Names and Classification Prosthetic groups, e.g. heme, are tightly bound to the apoenzyme. Copyright © Houghton Mifflin Company. All rights reserved. 3–51 16.14 Names and Classification Coenzymes are small organic molecules that are not tightly bound to the apoenzyme. They are often derived from vitamins. O O C C OH N Niacin, Vitamin B3 H H O C NH2 NH2 N N R R NAD+ NADH NAD+ and NADH are cofactors for many oxoreductases Copyright © Houghton Mifflin Company. All rights reserved. 3–52 16.14 Names and Classification Minerals are inorganic ions that act as cofactors. Common minerals: Zn2+ Mg2+ Copyright © Houghton Mifflin Company. All rights reserved. Mn2+ Fe2+ Cl1 3–53 16.15 How Enzymes Work In catalysis, the substrate fits into the active site of the enzyme, where it is held in place while the reaction occurs. E enzyme + S ES E substrate complex enzyme + P product This is called the enzyme-substrate complex, or ES complex. The reaction may then take place. Copyright © Houghton Mifflin Company. All rights reserved. 3–54 16.15 How Enzymes Work The active site is often a crevice-like region into which the substrate fits to form the ES complex. Copyright © Houghton Mifflin Company. All rights reserved. 3–55 16.15 How Enzymes Work The active site is often a crevice-like region into which the substrate fits to form the ES complex. The specificity of enzymes for substrates is attributed to the structures of active sites. Copyright © Houghton Mifflin Company. All rights reserved. 3–56 16.15 How Enzymes Work The “lock and key” model involves an active site with a fixed shape that accommodates only a certain substrate. Copyright © Houghton Mifflin Company. All rights reserved. 3–57 16.15 How Enzymes Work The “induced fit” model involves flexible active site that adapts to the structure of the substrate and binds to it. Copyright © Houghton Mifflin Company. All rights reserved. 3–58 16.16 Enzyme Activity Enzymes activity is the rate at which an enzyme converts substrate to product. “Turnover rate,” substrate molecules/minute or substrate molecules/second per molecule of enzyme “Turnover time,” seconds per molecule of product per molecule of enzyme Enzyme International Unit (EIU), concentration of enzyme that catalyzes 10–6 mole of substrate reaction per minute Copyright © Houghton Mifflin Company. All rights reserved. 3–59 16.16 Enzyme Activity Enzyme-catalyzed reactions are fast! Copyright © Houghton Mifflin Company. All rights reserved. 3–60 16.16 Enzyme Activity All chemical reaction rates increase with temperature. With enzymes, body temperature is optimal. At higher temperatures, the enzyme is denatured. Copyright © Houghton Mifflin Company. All rights reserved. 3–61 16.16 Enzyme Activity Acidity affects enzyme acitivity. Optimal pH for most enzymes is 7.0 to 7.5. Some digestive enzymes function best outside this range. Extremes in pH will denature the enzyme. Copyright © Houghton Mifflin Company. All rights reserved. 3–62 16.16 Enzyme Activity All chemical reaction rates increase with increases in reactant and catalyst concentration. If enzyme concentration is constant, rate will increase with substrate concentration to a maximum. Copyright © Houghton Mifflin Company. All rights reserved. 3–63 16.16 Enzyme Activity Reaction rate will increase if enzyme concentration is increased. Copyright © Houghton Mifflin Company. All rights reserved. 3–64 16.16 Enzyme Activity Enzyme activity can be reduced by inhibitors. Inhibition may be caused by a substance that occurs naturally in an organism to regulate enzyme activity. It may also be caused by a medicine or a poison. Enzyme inhibition can be irreversible or reversible. Copyright © Houghton Mifflin Company. All rights reserved. 3–65 16.16 Enzyme Activity Irreversible inhibitors form a covalent bond with a specific functional group of the enzyme and deactivate it. Cyanide binds to Fe3+ in cytochrome oxidase, so it can’t carry oxygen. Heavy metals bind to thiols. Cyt Fe3+ + O2 cytochrome oxidase Cyt Fe3+ CytFeO23+ reversible complex + CN1 CytFeCN2+ stable complex 1 + 2 CN S O 2 3 Copyright © Houghton Mifflin Company. All rights reserved. SCN1 + SO32 3–66 16.16 Enzyme Activity Some antibiotics inhibit bacterial enzymes. Penicillin inhibits a transpeptidase that is used in bacterial cell wall construction. Sulfa drugs interfere with synthesis of folic acid, which is required for growth of some bacteria. Folic acid is also essential to animals, but we get it in our diet. However, we need our intestinal bacteria! That’s why some antibiotics cause digestive distress. Copyright © Houghton Mifflin Company. All rights reserved. 3–67 16.16 Enzyme Activity Reversible inhibitors bind reversibly to enzymes. The enzyme-inhibitor complex is in equilibrium with its components. Enzyme + Inhibitor EI complex Shifting the equilibrium frees the enzyme. There are two types of reversible inhibition, competitive and noncompetitive. Copyright © Houghton Mifflin Company. All rights reserved. 3–68 16.16 Enzyme Activity Competitive inhibitors bind to the active site of an enzyme and block it. They compete with the substrate molecules for the active site. Noncompetitive inhibitors bind reversibly to an enzyme, but not at the active site. The inhibitor alters the shape of the active site. The enzyme cannot bind to the substrate. Feedback inhibitors are metabolic products that inhibit enzymes that catalyze their production. They can be competitive or noncompetitive. Copyright © Houghton Mifflin Company. All rights reserved. 3–69