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Function of Biomolecules
Department of Zoology
(3 Cr 30L + 30P)
and functions
What a lipid is and different types of lipids and their functions
Water
Carbohydrates
Lipids
Nucleic acids
Transport
Protection and defense Regulation of metabolic activities Means for movement, growth, and development Information storage
Each different type of protein has a characteristic amino acid composition and order Proteins range in size from a few amino acids to thousands of them
Folding is crucial to the function of a protein and is influenced largely by the sequence of amino acids
Protein Functions
1. Structural e.g. Collagen, elastin 2. Mobility e.g. Actin/myosin, tubulin, flagella 3. Receptors e.g. Insulin receptor 4. Ligands e.g. Insulin
Twenty amino acids used by the living organisms for synthesis of proteins
Charged R groups
The relationship between the chemical species and dissociation constant is expressed by the Henderson-Hasselbalch equation pH = pKa + log[A-]/[HA]
There are four levels of protein structure: primary, secondary, tertiary, and quaternary
The Four Levels of Protein Structure: Secondary Structure A proteins secondary structure consists of regular, repeated patterns in different regions in a polypeptide chain This shape is influenced primarily by hydrogen bonds arising from the amino acid sequence (the primary structure) The two common secondary structures are the alpha helix and the beta pleated sheet
The alpha helix is a right-handed coil The peptide backbone takes on a helical shape due to hydrogen bonds. The R groups point away from the peptide backbone and stabilize the structure by forming H bonds Fibrous structural proteins have a-helical secondary structures, such as the keratins found in hair, feathers, and hooves
The Four Levels of Protein Structure: Secondary Structure b pleated sheets form from peptide regions that lie parallel to each other Sometimes the parallel regions are in the same peptide, sometimes they are from different peptide strands This sheet like structure is stabilized by H bonds between N-H groups on one chain with the C=O group on the other
The primary determinant of the tertiary structure is the interaction between R groups
Hydrophobic side-chain aggregation and van der Waals forces, which help stabilize them
The ionic interactions between the positive and negative charges and hydrogen bonding between polar residues Disulfide bridges, which form between cysteine residues
Quaternary structure results from the ways in which multiple polypeptide subunits bind together and interact
This level of structure adds to the threedimensional shape of the finished protein
Hemoglobin is an example of such a protein; it has four subunits
Irregular contortions Secondary, Tertiary and from bondings Quaternary Structures between side Noncovalent Linkages chains. Hydrogen 4-20
van der Waals
Hydrophobic Hydrophobic clusters at the Ionic core of Covalent Linkages proteins Disulphide Bridges
The final conformation will be governed by the type of amino acids that make up the protein which will influence the folding pattern
Changes in amino acids can take place due to changes in DNA a process called mutation that can drastically change protein structure and therefore the function
Protein Denaturation
Changes in temperature, pH, urea, salt concentrations, and oxidation or reduction conditions can change the shape of proteins. This loss of a proteins normal three-dimensional structure is called denaturation.
Protein Modification
In some proteins further modification is needed for functioning
Glycosylation adding carbohydrate moieties which takes place in the golgi complex Adding lipid moieties especially in membrane proteins
Membrane Proteins
Lipid anchored proteins (a) Glycolipid covalent attachment by glycophosphatidyl inositol (GPI anchored proteins) (b) Covalent attachment of the protein to fatty acid like myristic acid or palmitic acid or the prenyl group (15-C franesyl hydrocarbons with repeating vinyl groups)
Protein Modification
In some proteins further modification is needed for functioning
Glycosylation adding carbohydrate moieties which takes place in the golgi complex Adding lipid moieties especially in membrane proteins
Covalent modification e.g. acetylation and methylation of Lys, methylation of Arg and His, phosphorylation of Ser, Thr or Tyr
Domains
The term domain is used to describe an area of a protein which is functionally or physically distinct Steroid Hormone Receptors
Inhibitory protein complex Transcription activating domain Hormone binding domain
Another example would be transmembrane proteins that have cytosolic, transmembrane and extracellular domains
Enzymes
Antibodies
Skin
Nails
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Fibrous proteins
Proteins which are folded to a more or less rod like shape
They consist of long fibers or large sheets tend to be mechanically strong are insoluble in water and dilute salt solutions play important structural roles in nature
Involved in structure: tendons ligaments blood clots, hair, hooves feathers etc., (e.g. Collagen, elastin, keratin and fibrin)
Fibrous Proteins
Keratin:
Ex: a-keratins
hair, wool, skin, and nails 3 a-helices held together by disulfide bonds Ex: b-keratins Feathers, scales large amounts of beta-pleated sheet structure
Fibrous Proteins
Collagen Connective tissue, skin, tendons, and cartilage
Consists of three polypeptide chains wrapped around each other in a ropelike twist to form a triple helix called tropocollagen; MW approx. 300,000 30% of amino acids in each chain are Pro and Hyp (hydroxyproline); hydroxylysine also occurs that contain OH groups for hydrogen bonding
The three strands are held together by hydrogen bonding involving hydroxyproline and hydroxylysine
With age, collagen helices become cross linked by covalent bonds formed between Lys and His residues Deficiency of Hyp results in fragile collagen
Globular Proteins
Proteins which are folded to a more or less spherical shape
They Tend to be soluble in water and salt solutions Most of their polar side chains are on the outside and interact with the aqueous environment by hydrogen bonding and iondipole interactions Most of their nonpolar side chains are buried inside Nearly all have substantial sections of a-helix and b-sheet
Prosthetic Group
Four N atoms from the porphyrin ring are attached to the Fe2+ center Fifth coordination site is occupied by a base (Histidine), of the globin protein Sixth coordination site can be occupied by O2 (oxyhemoglobin) H2O in (deoxyhemoglobin) CO in (carboxyhemoglobin)
Sickle-Cell Anemia
Results from a single mutation in the beta chain
Glu Val
(-) charge is changed to a nonpolar (hydrophobic) group This site of mutation is at the surface of the protein in the deoxy form of hemoglobin.
This results in the beta chains sticking together in the deoxy form
Immunoglobulins
The antibody molecule comprise of the immunoglobulin domain Immunoglobulin domain comprise of a 100 110 aa held together by intra-chain disulfide bonds that forms a compact loop within the chain (globular domain) 2 Heavy chains 2 Light chains
Functions of Antibodies
Enzymes lower G (Activation energy) but do not affect G (standard state free energy) for a reaction
Lect. 11-
Cofactors
In addition to the protein part, many enzymes also have a nonprotein part called a prosthetic group or a cofactor The protein part in such an enzyme is called an apoenzyme, and the combination of apoenzyme plus cofactor is called a holoenzyme. Only holoenzymes have biological activity; neither cofactor nor apoenzyme can catalyze reactions by themselves Cofactors form an intricate part of the active site and play a direct chemical role in the chemistry of the reaction
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Cofactors
A cofactor can be either an inorganic ion or an organic molecule, called a coenzyme
Many coenzymes are derived from vitamins, organic molecules that are dietary requirements for metabolism and/or growth
Nicotinamide adenine dinucleotide (NADH) Flavin adenine dinucleotide (FADH)
NH 2 N N N N R R S HO OH O R O
NADH
O P O O OH O P OHO R S
O R R OH
NH 2
O NH 2 N N N N R R HO O R S HO OH O OH OH O P O HO P O O R Me OH S N N O
Heme group
Metal atoms e.g. Zn++
FADH
Me
N NH
Classification of Enzymes
Class Reactions catalyzed
Hydrolases Lyases
Isomerases Ligases
hydrolysis
add/remove atoms to /from a double bond rearrange atoms combine molecules using ATP
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ES complex
E +
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Reaction Rate
Low
Temperature
High
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Increasing substrate concentration increases the rate of reaction (enzyme concentration is constant) Maximum activity reached when all of enzyme combines with substrate
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Factors Affecting Enzyme Action: pH Maximum activity at optimum pH R groups of amino acids have proper charge Tertiary structure of enzyme is correct Narrow range of activity Most lose activity in low or high pH
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Reaction Rate
11
pH
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Inactive Enzyme
Allosteric Regulation
Ca++ / Calmodulin
Proteolytic Cleavage
Some enzymes are produced as inactive Zymogens or proenzymes The active site of these enzymes are masked by a part of the molecule Cleavage of the masking portion by spontaneous degradation or other proteolytic enzymes leads to exposure of the active site and therefore activation
Some enzymes and enzyme complexes have fixed locations within the cells or body Nucleus: DNA replication, synthesis of tRNA and mRNA and some nuclear proteins Ribosomes: Protein synthesis Chloroplast: Photosynthesis Liver: Fatty acid metabolism, Gluconeogenesis, Glucose metabolism, Glycogen synthesis Adipose tissue: Fat metabolism
Enzyme Kinetics
For a given amount of enzyme the relationship between reaction velocity and substrate concentration
E + S k1 k -1 ES k2 P
V init =
Enzyme Kinetics
Lineweaver-Burk equation and plot allows us to determine Vmax and Km
Enzyme Inhibition
Cause a loss of catalytic activity There are FOUR types of enzyme inhibition: 1. Irreversible 2. Competitive 3. Non-Competitive 4. Uncompetitive
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Irreversible Inhibition
A compound interferes with the active site so as to disable it Commonly it is done by forming a stable covalent adduct with the enzyme
May also block substrate access to site Almost all are toxic substances
Competitive Inhibition
When an unreactive molecule bind to an enzymes active site and compete with the substrate to bind enzyme
Non-Competitive Inhibition
An inhibitor that binds to the enzyme, but not at the active site. In this case the inhibitor is not competing for the
active site
- Binding distorts the enzyme and reduces its activity e.g. allosteric regulation of the enzyme
Km remains unchanged
Uncompetitive inhibition
Substrate binding to enzyme is not inhibited Inhibitor binds to the ES complex occurs ESI complex is stabilized relative to ES complex so Km is reduced ESI complex is non-productive so Vmax is lowered
How enzymes aid in the catalytic process Bind substrates Lower the energy of the transition state Directly promote the catalytic event Either through acidic or basic side chains that promote addition or removal of protons Or through holding ions in correct position to participate in the catalysis Or by inducing stress that makes bonds labile Release the products