ENZYMES ESSAY QUESTIONS (ANSWERS) Question 1(a), (b)

RJC Prelims2005 P2/Q8

(a)

Describe the globular protein structure of an enzyme. [8] secondary structure of protein extensively folded up due to bond formation between side chains (R groups) to form tertiary protein with globular shape; Some enzymes are quaternary proteins consisting of 2 or more polypeptides held by weak/ non-covalent bonds formed between side chains; Disulphide bridges form between two cysteine monomers brought together by folding of the protein this is a strong bond that reinforces conformation; Ionic bonds form between a negatively charged acidic R group and a positively charged basic R group at certain pHs changing the pH will break these bonds; Hydrogen bonds form between polar side chains H that is part of OH or NH group becomes slightly positively charged and gets attracted to neighbouring electronegative O of a C=O group or N of an NH group; Hydrophobic interactions between hydrophobic R groups these interactions create a hydrophobic region within the molecule which excludes water; leaving the hydrophilic groups outside making the enzyme soluble; small region of the enzyme forms a precise 3-dimensional pocket or groove on the enzyme known as the active site; the active site typically consists of 3-12 amino acids; the rest of the polypeptide provides a framework that reinforces the configuration of the active site; some enzymes have other binding sites called allosteric sites on their surfaces which can bind to activator or inhibitor molecules;

(b)

Explain the effects of substrate concentration on the rate of enzyme-catalysed reactions. [4] At low substrate concentration, increase in substrate concentration results in an increase in the rate of enzyme-controlled reaction; Substrate concentration is the limiting factor at low substrate concentrations; As there is an increase in successful / effective collisions between enzyme and substrate molecules, leading to increase in enzyme-substrate complexes; At high substrate concentration, increase in substrate concentration results in no increase in the rate of enzyme-controlled reaction / constant rate of reaction at Vmax; All active sites are saturated with substrate molecules and any free substrate molecules will have to wait till the products to leave the active sites before binding to active site; Substrate concentration is no longer a limiting factor at high substrate concentration or enzyme concentration is now the limiting factor

[max 4m]

Question 1(c)
N2001 P3/Q9b

Explain the mode of action of enzymes in terms of specificity and activation energy.

[7]

collision theory-random collisions between substrate and enzyme; effective / successful collisions: binding of substrate at the enzyme active site in the correct orientation/precise position ; forming the enzyme-substrate (ES) complex; enzyme specificity: each enzyme can only bind to one type of substrate; substrate shape is complementary to enzyme active site shape; Lock and key hypothesis: active site has the precise configuration which the substrate fits exactly; Induce fit mechanism: active site not exactly complementary to substrate; binding of substrate induces a small conformational change to enzyme active site; temporary binding of substrate at enzyme binding site via ionic, hydrogen and hydrophobic / Van der Waals interactions; lowering of activation energy; holding substrate molecules in a precise position and orientation; binding of substrate at active site sets up physical stress in the substrate molecule; active site provides a favorable microenvironment for catalysis to occur; products formed do not fit active site, therefore are released from the active site; enzymes unaltered and can be used again;

Question 2 (a), (b)

Nov 2005 P2/Q8, RJC Prelims2005 P2/Q8c

(a)

Describe the effect of inhibitors on the rate of enzyme activity. [8] [for competitive inhibitors] inhibitor is similar in shape / structure with the substrate, it competes for and binds at / occupies the active site of the enzyme; thus blocking the substrate from binding with the enzyme; Number of enzyme-substrate complexes formed per unit time decreases: thus rate of reaction decreases; Effect of inhibition can be overcome by increasing substrate concentration; substrate molecules can compete effectively with the inhibitor molecules for enzyme active site and frequency of enzyme-substrate collisions increases; hence number of enzyme-substrate complexes formed per unit time increases reaction velocity being restored to the Vmax observed in the absence of inhibitor; [for non-competitive inhibitors] In non-competitive inhibition, Inhibitor has no structural similarity to the substrate; Binds to a site away from the active site on enzyme (allosteric); can alter globular structure of enzyme and rendering active site unable to bind substrate enzyme is still able to bind with the substrate at the active site but catalysis is unable to take place; Rate of reaction lowered as the effective enzyme concentration is lowered; Cannot be overcome by increase in substrate concentration and Vmax cannot be attained / restored; (decreases); As the substrate and inhibitor binds at different site on the enzymes;

(b)

Cells have metabolic pathways commonly made up of sequences of enzyme-catalysed reactions. Explain, with examples, the advantages of such sequences of enzyme-catalysed reactions. [6] *Must quote a relevant example In the production of isoleucine from threonine; the build up of isoleucine (end product); will inhibit threonine deaminase (first enzyme of the anabolic pathway) and prevent further production of isoleucine; Prevent excess maunfuacturing of products; Ensure reactants are used more efficiently; Products becomes the substrates of next reaction; Each reaction catalysed by a different enzymes; Intermediates fomed can be fed into different pathways;