Biology objectives 1

(a) Describe and interpret drawings and photographs of typical animal and plant cells as seen under the electron
microscope.
(b) Outline the functions of the membrane systems and organelles.
Membrane Systems
a. Within cell
• Compartmentalisation, allows division of labour / specialisation or any appropriate example of different
organelles serving different functions
• Synthesis of lipids in sER
• modification and packaging of proteins in Golgi apparatus any other relevant example
• Localisation of enzymes and substrates (ie in one place), helps increase efficiency of reactions by allowing
the concentration of specific substances within parts of cells
• E.g. enzymes and substrates required for Krebs cycle are compartmentalised in the mitochondrial matrix
• Allows the creation/maintenance of special environment eg. pH for optimal reactions
• Regulates passages of substances in and out of organelles
• Increase surface area for attachment of more enzymes or electron carriers -- inner membrane of
mitochondria that is greatly folded to form cristae
• for the attachment of more ATPase involved in ATP production
• provides a platform for the attachment of hydrogen / electron carriers that requires a specific spatial
arrangement E.g. linear arrangement of hydrogen / electron carriers of the electron transport chain on the
inner mitochondrial membrane facilitates the transfer of hydrogen / electrons down the chain
• Intracellular transport e.g. rER  GA  secretory vesicles
• Protection, compartmentalisation of hydrolytic enzymes into lysosomes, prevents the digestion of cellular
contents

b On the surface of cells

• The phospholipid bilayer/plasma membrane of the plasma membrane creates a boundary between the
contents of the cell and its external environment / delimits the cell from the environment;
• allowing it to function as a separate entity from the external environment;
• Partially or selectively permeable membrane regulates the passage of substances in an out of the cell;
• Allows the entry of nutrients e.g. entry of glucose/ amino acids);
• exit of waste products (e.g. exit of carbon dioxide)
• Via simple and facilitated diffusion, osmosis; bulk transport; active transport; Na+/K+ pump;
• Phospholipids – fat-soluble substances;
• Proteins – polar/hydrophilic/charged molecules;
• Helps to maintain constant internal environment within the cell;
• Receptor proteins enable the cell to communicate with external environment; sense changes and respond;
• Receptor proteins receive chemical messenger molecules
• Glycoproteins and glycolipids/ carbohydrates attached to the proteins and/or phospholipids enable cell-to-
cell recognition
• immune system for recognition of foreign pathogens, helps to maintain structural relationships with
neighbouring cells, enable cell-to-cell adhesion, enables tissue formation

Nucleus
Largest organelle within the eukaryotic cell; spherical; diameter of 10-20µm
Nuclear envelope
o Double membrane
o outer membrane continuous with endoplasmic reticulum
o nuclear pores  places where the 2 membranes fused; function as channels whereby molecules can
 move between the nucleus and cytoplasm.
Nucleoplasm -- Semi-fluid matrix that fills the nucleus
Chromosome / Chromatin
o Most of a cell’s DNA located in the nucleus and is organized along with proteins (histones) into
chromosomes.
o Chromatin - highly elongated threads; during nuclear division, the chromatin becomes visible as
chromosomes.
o 2 types of chromatin- Heterochromatin (highly condensed DNA, genes are inactive) and Euchromatin (not
so tightly packed, genes are active)
Nucleolus
o Most visible structure (within a non-dividing nucleus)
o One or more nucleoli may be found within the nucleoplasm.
o Nucleoli are responsible for the synthesis of ribosomal ribonucleic acid (rRNA), which is a component of
ribosomes.
• Contains hereditary material (chromosomes) of an organism.
• Essential for cell division
• Controls and directs activities of a cell by regulating protein synthesis. DNA within nucleus contains genes
which are templates used in protein synthesis (e.g. enzymes)

Mitochondria
Cylindrical or rod-shaped.
Vary in size (width ranges from 0.5-1.5µm; length ranges from 3-10µm).
Bound by double membrane: outer and inner membrane separated by inter-membranal space
Outer membrane: smooth continuous boundary.
Inner membrane:
o extensively folded  forming cristae which project into the semi-fluid matrix. (Increase SA of contact)
o for attachment of photosystems for efficient capture of light energy
o site for attachment of electron transport system, these can be attached in an ordered sequence for
improved efficiency.
o Attachment for enzymes such as ATP synthase involved in production of ATP
o impermeable to H+ thereby allowing the buildup of high H+ concentration in the intermembranal space
Matrix contains circular DNA and 70S ribosomes, which are present in prokaryotic cells.
Presence of DNA  mitochondrion partially independent of control of nucleus while 70S ribosomes  protein
synthesis occur
• Responsible for cellular respiration, a series of biochemical reactions that result in formation of ATP
(Oxidative Phosphorylation). A metabolically active cell may have more than 1000 mitochondria.

Chloroplast
Large organelles (diameter: few µm; length: 5-10µm)
Bound by double membrane: outer and inner membrane.
Outer membrane: smooth continuous boundary.
Inner membrane highly folded to form
o thylakoids (or lamellae)
o thylakoids are stacked to form grana (singular: granum)
o Stacks of grana are joined by intergranal lamellae.
o Both grana and integranal lamellae contain photosynthetic pigments (e.g. chlorophyll).
Interior of chloroplast is filled with gel-like matrix called stroma. Stroma contains circular DNA and 70S ribsomes.
May also contain starch grains.
• Chloroplasts are sites of photosynthesis. The light-dependent reactions of photosynthesis occur on the
lamellae while the light-independent reactions occur in the stroma.
• Synthesis of ATP through photophosphorylation.

Endoplasmic reticulum
Two types: Rough endoplasmic reticulum (rER) and Smooth endoplasmic reticulum (sER)
rER: Originates from outer membrane of nuclear envelope.
• Consists of a network of flattened membrane-bound sacs called cisternae (singular: cisterna) with
ribosomes attached on the outer surface.
• In sections, it typically appears as pairs of parallel lines (membranes) running through the cytoplasm.
sER: consists of a series of interconnected tubules; lacks ribosomes.
Although the two types of ER can be distinguished and each has its distinct functions, their membranes are
connected and their internal spaces are continuous.
• Rough ER:
o site where proteins that are meant to be exported or kept in a vesicle are synthesized (by the attached
ribosomes) and transported. Proteins are transported either through cisternae to their destinations or
packaged in vesicles and dispatched to other parts of the cell or secreted across cell surface membrane
(via exocytosis),
o folding of protein into specific shape and
o Modification of proteins (e.g. phosphorylation, attachment of carbohydrate chains  glycoprotein)
• Smooth ER:
o site of synthesis of lipids,
o metabolism of carbohydrates,
o detoxification of drugs and poisons
o stores calcium ions required for contraction in muscle cells

Golgi apparatus
Consists of a stack of flattened membrane-bound sacs also known as cisternae and a system of associated vesicles
called Golgi vesicles.
Cisternae are continually being formed at one end of the Golgi body (i.e. “forming” face or cis face) and continually
being broken down into vesicles at the other end (i.e. “maturing” face or trans face).
A vesicle that buds from ER adds its membrane and contents of its lumen to the Golgi body by fusing with the
“forming” face of the Golgi body.
• Chemically modifies the carbohydrate portions of the glycoprotein. (Carbohydrates are first added on in the
ER.)
It can also synthesise some carbohydrates.
• Sorts and packages proteins (and chemically modifies them if necessary) before transporting them to
various cellular location.
Eg. Some Golgi vesicles carry membrane proteins from the “maturing” face of Golgi body to cell surface
membrane, fusing with it and enlarging it. Other Golgi vesicles carry secretory proteins (e.g. enzymes) to the cell
surface membrane for release to the exterior (exocytosis).
• Formation of lysosomes.
• Involved in the formation of new cell wall. After nuclear division, Golgi vesicles move to the region between
two newly formed daughter nuclei. Their membranes fuse and become the new cell surface membranes of the
daughter cells, while their contents (e.g. pectins) contribute to the middle lamella and cell wall matrix.

Lysosomes
Small spherical vesicles (0.2-0.5 µm) formed from Golgi body.
 consist of hydrolytic enzymes (e.g. lipases, proteases, nucleases) surrounded by a single membrane.
Contents are acidic and enzymes have a low optimum pH.
• Fuse with vesicles formed by endocytosis and release hydrolytic enzymes into the vesicles to digest the
material within. The material may be taken in as food (e.g. Amoeba), or for defence of body (e.g. white blood cell
engulfing bacteria). The products of digestion are absorbed and assimilated by cytoplasm of cell.
• Engulf and digest worn-out organelles within a cell (autophagy)
• Self-digestion of a cell by releasing the contents of lysosomes within the cell (autolysis). Occurs after cell
damage or cell death.

Ribosomes
Non-membrane bound organelle
Composed of proteins and ribosomal RNA (rRNA) in roughly equal quantities.
Can be found attached to the endoplasmic reticulum or freely suspended in the cytosol.
An eukaryotic ribosome has a diameter of about 20nm.
Has a sedimentation coefficient of 80S and consists of two subunits: a small subunit of 40S and a big subunit of
60S. The sedimentation coefficient of a particle depends on its molecular size and geometrical shape. (prokaryote:
70S – 30S and 50S)
Ribosomes normally occur in clusters called polyribosome or polysomes. This allows for the simultaneous synthesis
of many polypeptide chains from a single mRNA.
Small subunit allows attachment of mRNA to form initiation complex
Large sub-unit consists of 3 binding sites: mRNA binding site, 2 tRNA binding sites
• Site of protein synthesis (translation) where the amino acids are joined together to form polypeptide chain
via the formation of peptide bonds.
Specifically:
o Provides the platform for translation through the P site (holds the first aa-tRNA complex) and A sites
(receives aminoacyl-tRNA complex)
o Freely-floating ribosomes in cytosol produces protein that function within the cytosol.
o Ribosomes attached to rER synthesize proteins that are meant for insertion in to the membrane and for
packaging within certain organelles.
• Cells active in protein synthesis have a large number of ribosomes.

Centrioles
exists as a pair of rod-like structure positioned at right angles to each other and situated next to the nucleus (at a
region known as centrosome).
Size: diameter: 200nm; length: 500nm
Found in both animal cells and lower plant cells but are absent in higher plant cells.
Transverse section of a centriole shows 9 triplets of microtubules arranged in a ring. Each triplet attached to each
other by fibrils.
Microtubules are long hollow tubes made up of tubulins.
• Acts as microtubule organizing centres (MTOC) for the formation of the spindle fibres during mitosis and
meiosis.

(c) Describe the formation and breakage of a glycosidic bond

Bond formed by condensation, involving the removal of water from 2 monosaccharides
• 1.4 glycosidic bond formed between the carbon atom 1 of one monosaccharide and carbon 4 of the other
(found in starch and glycogen – gives it a helical structure so that is it compact and takes up little space in cell)
• 1,6 glycosidic bond formed between carbon atom 1 and carbon atom 6 of another. (found in starch and
more in glycogen – allows molecule to be highly branched which increases surface area for enzymes to act on it
at he same time to convert it to glucose)
• 1,4 beta glycosidic bond formed in cellulose, where every alternate residue is rotated 180° so that –OH of
carbon 1 comes alongside –OH of carbon 4. Forms a straight chain with –OH groups projecting outwards in all
directions, allowing interchain hydrogen bonding between neighbouring chains to be most effective since surface
area for contact is maximised
Bond is broken through hydrolysis, involving the addition of water to the polysaccharide.
• Chemical method – incubate disaccharide with dilute acid at 100°C
• Enzymatic method – incubate with enzyme e.g.sucrase, maltase, lactase or amylase for starch

(d) Analyse the molecular structure of a triglyceride and a phospholipid and relate these structures to their
functions in living organisms
Similarities
• Both are lipids made up of C, H and O.
• Both contain glycerol and fatty acids are attached to glycerol via an ester bond
• A condensation reaction results in the formation of water.
• Hydrocarbon lengths may differ and degree of saturation also differs
Phospholipids
• Compound lipid with 2 fatty acids and a phosphate group that is joined to one of the hydroxyl groups of the
glycerol resulting in a phosphodiester bond.
• Consists of hydrophilic phosphate head facing the aqueous environment and 2 hydrophobic fatty acid
carbon tails facing the internal core.
• Being amphiphatic, serves as part of the membrane structure and helps to form a boundary that enables
cell to function as an entity separated from the environment, allows formation of organelles within the cell, allows
diffusion of lipid soluble molecules across the membrane down their concentration gradient – regulate the
movement of substances in and out of the cell (selective permeability)
• Held together by hydrophobic interactions-- giving the membrane stability
• Molecules are free to move laterally within the membrane – provides fluidity for formation of vesicles during
endocytosis and exocytosis
Triglycerides
• Simple lipid with 3 fatty acids joined to a glycerol, making it hydrophobic
• Large and uncharged molecules, thus insoluble in water. It can be stored in large amounts without any
effect on osomotic potential and is prevented from diffusing out of the cell -- Serves as a storage form of energy
• Provides more energy than carbohydrates as it contains more carbon atoms per gram. (38kJ/g compared
to 17kJ/g for carbohydrates)
• Releases twice as much metabolic water than carbo when oxidized due to greater hydrogen atoms
• Good thermal insulator and prevents excessive heat loss
• long term energy store in hibernating animals
• gives buoyancy to aquatic mammals as it is less dense than water
• are components of the myelin sheath as it acts as an electrical insulator.

(e) Describe the structure of an amino acid and the formation and breakage of a peptide bond.
Amino acid has an amino group, a carboxyl group, a hydrogen atom and a variable R group that are attached to a
central a-carbon (except for glycine). At physiological pH, both the carboxyl and amino groups are ionized and form
a zwitterions. R group carries acidic, basic, polar and non-polar properties. Amino acids are colourless, crystalline
solids which are generally soluble in water but insoluble in organic solvents. In solution, they can act as buffers as –
COO groups can pick up additional H+ added while NH3+ can donate H+ to neutralize the small amounts of OH-
A peptide bond is formed through condensation of 2 amino acids. It forms between the OH of the carboxyl group
and the NH of the amino group of the other amino acid
Peptide bond in broken via hydrolysis.

(f) Explain the meaning of :

Primary structure - type, number and sequence of amino acids in chain, variable group in an amino acid is the R
group. It determines the properties and shape as well as biological functions of polypeptide.
Secondary structure – localized folds and coils that occur as a result of hydrogen bond formation at regular intervals
along the polypeptide backbone between amino acids that are lying close to each other.
1. alpha-helix
the backbone winds around the long axis, resulting in a right handed-coil with the hydrogen bonds all
aligned parallel to the axis and all the side groups projected outwards. There is a repetitive sequence of
amino acids and the helix is stabilized by intrachain hydrogen bonds between the nth carbonyl group to
(n+4) amino group of the peptide bond. There are 3.6 aa residues in one complete turn of the helix.
2. beta pleat
2 or more regions of the polypeptide chain lying parallel to each other associate together and hydrogen
bonds are formed between the carbonyl group of 1 chain and the amino group of adjacent chains. R
groups project above or below the pleated sheet
Tertiary structure – polypeptide with a unique sequenceof aa folds extensively upon itself to form a precise, compact
globular shape; Interactions involve R groups of amino acids that are far away; Resulting in the formation of a
specific and compact 3D conformation or shape; Formation of Active sites for enzymes that are complementary to
the substrates; Hydrophilic groups are exposed but hydrophobic groups found inside the core – makes the protein
soluble (NB: solubility is important for transportation; for reactions to take place). 4 types of bonds bet the R groups
Quaternary structure – the association of 2 or more polypeptide chains by hydrophobic interactions, hydrogen and
ionic interactions to form a precise structure.
Describe the type of bondings which hold proteins in shape
Hydrogen – form when a hydrogen atom that is covalently bonded to one electronegative atom is simultaneously
attached to another electronegative atom
Ionic – a charged R group can attract an oppositely charged R group in another part of the chain. Electrostatic
attraction is very much pH dependent
Disulfide – formed between the side chains of 2 cysteine residues.
Hydrophobic interactions – non polar R groups tend to cluster together in water, where hydrophobic R groups being
present in the interior, whereas hydrophilic R groups tend to be on the outside to attract water. This is a major force
driving proper protein folding.
These bonds are important in maintaining the 3D shape of the protein;
If bonds are disrupted, the protein will unravel;
Shape is not maintained means loss of function; Shape is important for the functioning of the protein

(g) Analyse the molecular structure of a dimeric enzyme with a quaternary structure e.g. HIV protease and
collagen and relate these structures to their functions

Features Fibrous protein Globular protein

1) Named example Collagen HIV Protease
2) Overall level of Quaternary Quaternary
organisation
3) Components/ - Many subunits (tropocollagen) Made up of 2 subunits, with 99 aa in each.
Subunits - each subunit consists of 3 polypeptides - Each subunit is made up of 1 polypeptide,
(Both are made up of - each subunit exhibits quaternary level (triple with 2 beta pleated sheets and a small
subunits) helix) alpha helix.
- within each subunit – there are 3 polypeptides - each subunit exhibits a tertiary level of
(It is impt to clearly each showing a secondary level of organisation organisation
differentiate between - Interaction within each subunit is due to (2 subunits held together to form a
interactions that are hydrogen bonding quaternary structure)
within the subunit - Interaction between subunits is due to
with those that are covalent bonds - Interactions within each subunit are
between subunits.) - Polypeptide chains (within each subunit) are hydrogen bonds, ionic bonds and
cross-linked at intervals to form long fibres. hydrophobic interactions
- For each polypeptide, the amino acid - interaction between subunits is due to non-
sequence is remarkably regular. For example, covalent bonds
almost every third amino acid in the polypeptide - Polypeptide chain(within each subunit) is
chain is glycine. Glycine is a helix breaker thus tightly folded to form a spherical shape;
helix has a slightly kinked conformation that - For each polypeptide, it does not exhibit
allows it to wound round tightly around each regularities in the amino acid sequence;
other

4) Variation in aa Amino acid sequence may vary slightly Amino acid sequence is highly specific and
sequence bet 2 of the between two samples never varies between two samples;
same protein
b) Length of Length of polypeptide chain may vary in two Length of polypeptide chain is always
polypeptide samples; identical in two samples;
5)Further Subunits (Tropocollagen) -> covalent bonds 2 identical subunits giving rise to a functional
organisation and staggered arrangement(ensures that there HIV protease. Active site formed at the
are no weak areas) give rise to fibrils -> fibrils interface of 2 subunits and consists of 2
bundled together to give rise to fibres aspartic residues. Beta sheets help to form
(which happens outside the cell) the flaps of the active site, that holds the
substrate within the active site
6) Stability (to Stable More pH and temperature sensitive as there
changes in pH and Due to the high number of cross-linkages that are many non-covalent bonds that important
temp) in ensuring that the enzyme has a specific 3D
are formed between the polypeptides and conformation
between tropocollagen

7) Solubility in water Insoluble in water; Dissolve in water to form colloidal solutions;

Due to large number of hydrophobic R groups Due to hydrophilic R groups of amino acid
of amino acid residues on the exterior of the residues jutting outwards from their
molecules; molecules. Hydrophobic groups kept within
the molecule.
(Insolubility is important to collagen as it serves (Solubility is important to HIV protease as it
as a structural protein) acts as a hydrolase and chemical reactions
occurs in an aqueous medium)

8)most important Secondary. Allows a large surface area for
level of organisation cross-linking so as to increase tensile strength
9) Function Structural and supportive function - eg. tendon,
bone, teeth. Main protein of connective tissue in
animals and creats the body’s physical
structure. Also gives skin strength and elasticity.
Tertiary. Allows specific 3D conformation of
active site to be formed so that only substrate
complementary to it will be acted upon
Metabolic function - Cleaves the viral
polyprotein into smaller functional protein
chains, is subsequently assembled to form
new viral coat structure

Other similarities:
1) both made up of amino acids (as monomers)
2) the amino acids are joined together by peptide bonds (formed through condensation reaction)
(h) Explain the mode of action of enzymes in terms of an active site, enzyme substrate complex, lowering the
activation energy and enzyme specificity
Active sites make up a small portion of the enzyme molecule (about 3 to 12 aa residues)
Enzymes decrease activation energy by
- Colliding with substrate at the correct orientation, causing the substrate molecules to bind to the
enzyme molecule at its active site, forming E-S complex where the chances of reaction occurring
is greatly enhanced as enzyme hold substrate in correct orientation. so that the substrates can
react together, then products are released
- bond break more easily through physical stress that is put on it when substrate bins to active site
- R groups at active site change charge on substrate when put very close to each other altering the
distribution of electrons to increase reactivity of substrate
Enzyme specificity
Lock and Key Hypothesis – shape of substrate is complementary to shape of active site of enzyme. Substrate is the
key to unlock the enzyme. Once products are formed, they no longer fit into AS, leaving the active site free to
receive further substrate molecules
Induced Fit hypothesis – binding of substrate to AS enables a conformational change in shape of enzyme, which
enables the substrate to fit more snugly into the AS. AS has a complementary site only after substrate is bonded to
it which enables enzyme to perform its catalytic function more effectively.

(i) Follow the time course of an enzyme –catalysed reaction by measuring rates of formation of products
e.g. using catalase or rate of disappearance of substrate e.g. using amylase.
Signal – changes with either enzyme or substrate concentration. E.g. Colour changes, gas produced, mass
changes, pH changes and volume changes.
Initial rate measurement is repeated under different conditions and a graph of rate vs the factor is plotted. Each
point is taken from a separated initial rate measurement (or better still is an average of several initial rate
measurements under the same conditions)

(j) Investigate and explain the effects of temperature, pH, enzyme concentration and substrate concentration on
the rate of enzyme catalysed reactions and examine these effects.
Denaturation refers to the disruption of the characteristic 3D arrangement of polypeptide chain(s) due to the
unfolding of the chain(s) resulting in loss of biological activity.
Temperature
Rate of reaction increase with increasing temperature until optimal temp is reached. For most enzymes, the rate of
reaction is doubled for every increase of 10°C.
As temp increase, increase in KE of E and S molecules results in increase in number of effective collision between
E and S per unit time. More E-S complexes form and thus rate of rxn increase
When tem goes beyond optimum, rate decreases rapidly as enzyme denatures. Excessive heat disrupts ionic and
hydrogen bonds which stabilize secondary and tertiary structures of the enzyme, thus causing enzyme to unfold and
precise shape of active site is lost.

pH
At optimum pH, the intramolecular bonds which stabilize secondary and tertiary structures of the enzyme are intact.
The conformation of the active site is most ideal for binding the substrate and frequency of successful collisions
between enzyme and S is the highest. Hence, more E-S complexes form and thus rate of rxn increase.
At pH lower of higher than optimum, concentration of H+ ions changes and this alters charges of R groups of amino
acids residues on the enzyme. Ionic bonds that maintain conformation of enzyme would be disrupted and biding of
substrate is affected
 Substrate concentration
At low (S), increasing (S), rate of reaction increases as a lot of free active sites are available. Any S present will be
immediately catalyzed. Rate of reaction is thus proportional to increase in (S). With a further increase in (S), rate of
reaction increases at a slower rate as more active sites are occupied. Number of free AS decreases. S molecules
have to compete for active sites as the probability of finding a free AS decreases. Further in crease in (S), rate of
reaction remains constant as all AS are used up at any one time, enzymes are fully saturated. Any S will have to
wait for products to leave enzyme before it can complex. Enzymes are thus working at their maximum rate.

Enzyme concentration
At low (E), amount of enzyme is limiting, all the AS of the enzyme are saturated at any one time. Increase in (E)
results in a proportional increase in the rate of reaction as more AS sites are available for the formation of E-S
complex at any one times. At high (E), further increase in enzyme does not increase rate of reaction. Graph
plateaus off due to insufficient substrate molecules competing for available AS. The amount of substrate of another
factor becomes limiting. Adding extra substrate can result in increase in rate of reaction then.

(k) Explain the effects of competitive and non-competitive inhibitors on the rate of enzyme activity
Features Competitive inhibition Non - competitive inhibition
structure resembles the substrate in terms of shape bears no structural similarity to the substrate
molecule
Binding binds reversibly to the active site of the enzyme binds to inhibitor site and not the active site of
site and the enzyme, Binding of inhibitor may be
reversibility reversible (or irreversible)
Effect of competing with the substrate for binding at the active altering the globular / 3D structure of the enzyme
inhibitor site of the enzyme, preventing formation of ES and the configuration of the active site, thus
binding complex rendering active site unreceptive to the substrate

Effect of Effect of inhibition can be overcome by increasing Effect of inhibition cannot be overcome by
increasing substrate concentration, as high (S) increases the increasing substrate concentration
substrate chance of effective collisions between substrate and
con. enzyme over that between inhibitor and enzyme
Effect on At high substrate concentration, rate of reaction can Inhibitor decreases the Vmax of the reaction
Vmax and reach the same Vmax as the non-inhibited reaction Inhibitor has no effect on the Km since they do
Kmax Inhibitor increases the Km for a given substrate – i.e.
not compete with the substrate for the active site
in the presence of the inhibitor, more substrate is
of the enzyme
needed to achieve Vmax / because inhibitor lowers
(1/Km is not affected – affinity of enzyme for
affinity of enzyme active site for the substrate
substrate)
• End-product inhibition:
i. Accumulation of end product of a metabolic pathway which involves a series of enzyme-catalyzed
reactions may have an inhibitive effect usually on the enzyme which catalyzes the first reaction in the
pathway
ii. Inhibitive effect is self-regulatory / if the end product is used up, inhibitive effect is removed

• Allosteric inhibition:
i. Inhibitor binds to allosteric site and not the active site of the enzyme
ii. Binding of inhibitor is reversible
iii. Inhibitor regulates the activity of the enzyme when bound, it alters the configuration of the enzyme active
site, rendering active site unreceptive to the substrate. When unbound, configuration of active site returns
to the original / enzyme regains its productivity

(l) Explain the importance of mitosis in growth, repair and sexual reproduction
• Mitosis ensures that the 2 daughter cells contain genetically identical sets of chromosomes as the parent
 cell
• Important for growth of multicellular organism – after the human sperm fertilizes the ovum, the resultant
zygote cell divides multiple times via mitosis to develop and eventually grow into a multicellular adult.
• Important for repair of worn-out parts of the body – damaged or old cells can be replaced with new,
genetically identical cells
• Important for the bases of asexual reproduction(the production of offspring form a single organism, without
the formation and fusion of gametes like sperm and ovum)
- when a unicellular organism like the amoeba divides to form identical offspring, division of one cell
reproduces an entire organism
- mitosis on a larger scale is involved in vegetative propagation of certain plants that grow from
buds eg. Potato, ginger and onions
- gives advantage to species that has successfully colonized a particular habitat, helps to quickly
establish a colony of individuals that are genetically identical to the parent, if not, offspring may
not be able to adapt to the habitat and die.

(m) Explain the need for the production of genetically identical cells and fine control of replication
• mitosis helps to maintain genetic stability by forming daughter nuclei that have the same number of
chromosomes and same genetic makeup as the parent.
- Chromosomes are duplicated before mitosis begins and replication is semi-conservative and
makes use of complementary base pairing and thus is very accurate
- Arrangement of chromosomes on the spindle during metaphase ensures that chromosomes are
shared equally between the 2 daughter nuclei. Separation of sister chromatids at anaphase so
there is no variation in daughter nuclei.
• Cell division is finely controlled by multiple cell cycle checkpoints that verify whether the processes at each
phase of the cell cycle have been accurately completed before progression in to the next phase. A complex
network of proteins ensures that these events occur at the proper times. Intracellular and extracellular
signals block cell-cycle progression at checkpoints if certain events have not yet been completed. After the
restriction point, the cell is committed to replicating its genome and dividing, completing one round of the
cell cycle. If, prior to the restriction point, cells sense inadequate growth conditions or receive inhibitory
signals from other cells, they enter G0 (G-zero) phase. In the G0 phase, they are maintained for prolonged
periods in a nondividing state. Whereas some conditions cause cells to enter the G0 phase, others trigger
apoptosis . One such signal that may trigger apoptosis is if a cell's DNA has undergone significant
damage.
• G1/S checkpoint
- replication of DNA will not occur in the absence of growth factors and before minimum cell size is
reached. During this phase, cells monitor their environment and determine if conditions, including
the availability of nutrients, growth factors and hormones, justify DNA replication.
• G2 checkpoint
- Mitosis is prevented if DNA is not replicated properly. Should the cell enter M phase even when
not all genes have been replicated, some genes would be missing in the daughter cell.
- important mechanisms that control the completion and fidelity of DNA replication and that prepare
the cell for entry into mitosis occur here
• M checkpoint / Spindle attachment checkpoint
- Chromosome separation is delayed if the chromosomes are not properly attached to the mitotic
spindle. Should the cell transit into anaphase when not all chromosome are aligned properly at
metaphase, aneuploidy or polyploidy will result.
- cyclins and regulators prevent cell cycle from continuing if there is defect in the DNA. The
ubiquitin protein ligase complex called anaphase-promoting complex (APC) is then responsible
 for initiating Anaphase

(n) Explain how uncontrolled cell division can result in cancer and identify factors which can increase the chances
of cancerous growth
Cancer is the consequence of dysregulation of checkpoints of cell division resulting in uncontrolled cell division.
Most cancers originate in a single cell which undergoes several mutations either due to gain of function mutations of
proto-oncogenes or loss of function mutations of tumour suppressor genes which accumulate during cell division
until it acquires the ability to break free from the cell cycle controls.
Cancerous cell divides uncontrollably and at a higher rate than normal cells until a tumour is formed. The tumour
may be malignant where angiogenesis (growth of a new network of blood vessels) occurs and the cell goes through
metastasis and is carried to other parts of the body via the circulatory or lymphatic system which may result in new
cancers in other parts of the body. Cancerous cells do not undergo apoptosis and can divide indefinitely due to
presence of telomerase; they do not show contact inhibition and do not stop dividing further when in contact with
other cells, they fail to differentiate properly to become specialized tissues.
Factors that increase the chances of cancerous growth
• Internal factors
- Genetic e.g. hereditary disposition such as the inheritance of one copy of the BRAC1 gene greatly
increases the lifetime risk of breast cancer by 70%
- Chances of Cancer-causing mutations occurring and accumulating increases with Age
- Only females can have breast cancers
• Chemicals
- tar in cigarette smoke and ethidium bromide either damage or alter DNA
• Exposure to radioactivity
- excessive UVB light is largely responsible for carcinogenic properties of sunlight. Mutations in the
p53 gene is triggered by this
- ionizing radiation like X-rays can penetrate the nucleus and form the damaging ions inside cells
that can break or mutate DNA
• Viruses
- retroviruses contain a gene that alters the host cell division genes. When the viral genome inserts
hear the cellular proto-oncogene, it will now be under the control of the viral promoter, switching
the genes on and causing them to become oncogenes. (gain of function)
- When the viral genome inserts into a particular site in the host chromosome, it may disrupt a
tumour suppressor gene of the host cell. Viral infection may thus increase the chance of getting
cancer by causing loss of function mutation in TSG
-
(o) Describe the behaviour of chromosomes during the mitotic cell cycle and the associated behaviour of the
nuclear envelope, cell membrane and centrioles
Prophase – condensation of chromatin to from chromosomes. Centrioles move to opposite ends of the cell and
asters develop from each pair. Nucleolus and nuclear envelope disintegrate. Microtubules arise from asters and
form a spindle.
Metaphase – chromosomes arrange themselves at the equator of the spindle and become attached to spindle fibres
at their centromeres. The outer kinetochore interacts with the spindle fibres, while the inner kinetochore is tightly
associated with the centromere.
Anaphase - centromere splits. Sister chromatids of each chromosome separate and move in a V-shape towards
opposite poles of the spindle.
Telophase – separated sister chromatids reach their respective poles and become chromosomes of daughter cells.
Chromosome decondense and become chromatin. Spindle fibres disintegrate and nuclear envelope and nucleolus
reform.
Cytokinesis in animals – a furrow develops in the cell membrane on either side of the cell and deepens. Eventually,
the cell membrane in the furrows join up and completely separates the 2 daughter nuclei and forms 2 daughter cells
Cytokinesis in plants – a series of Golgi apparatus form and line up in the middle of the parent cell. They fuse to
form the cell plate which extends outwards across the equator of the parent cell. They make up the cell walls of the
daughter cells and its membrane makes up the cell surface membrane of the daughter cell. Cell plate eventually
fuse with parent cell wall and cell membrane, thus separating the 2 daughter nuclei.

(p) Explain what is meant by homologous chromosomes

• chromosomes that occur in pairs
• 1 paternal and the other maternal
• same morphology eg. size and length, position of centromere
• same linear sequence of genes
• may contain different alleles
• pair with each other during prophase 1, meiosis.

(q) Describe the behaviour of chromosome during meiosis and the associated behaviour of the nuclear envelope,
cell membrane and centrioles.
Meiosis = reduction division
Interphase 1 (S phase)- DNA replicates. Each chromosome thus consists of 2 identical chromatids
Prophase 1 - Chromosomes shorten/thicken/coil up/condense. Nuclear envelope and nucleolus disintegrate.
Centroles migrate to opposite poles and spindle tubules form. Pairing of homologous chromosomes and formation
of chiasmata where crossing over takes place (exchange of genetic material)
Metaphase 1 - Bivalents are orientated randomly on the equator, allowing the daughter cells to inherit random
mixture of paternal and maternal chromosomes. Total number of possible chromosome combinations is 2 to the
power of n (haploid number of chromosomes)
Anaphase 1 - Whole chromosome is pulled apart and spindle fibres shorten to aid in this process. 2 centromeres of
each bivalent remain intact. Bivalents move towards respective poles in a V shape
Telophase 1 – chromosomes reach their respective opposite poles and each pole will have a haploid set of
chromosomes. Nuclear envelope and nucleolus reform.
Cytokinesis – cytoplasmic cleavage resulting in the physical separation of 2 haplios nuclei into 2 daughter cells.
(no interphase)
Prophase 2 –Nuclear envelope and nucleolus disintegrate. Centroles migrate to opposite poles and asters form
spindle fibres.
Metaphase 2 – chromosomes line up along the equator at right angles to the axis at metaphase 1 randomly.
Anaphase 2 – centromeres split and are pulled apart with aid of spindle fibres, sister chromatids move to opposite
poles in a V shape.
Telophase 2 – chromosomes decondense and nuclear envelope and nucleolus reform. 4 haploid nuclei which hare
genetically different from each other is produced.
Cytokinesis – separation of nuclei into 2 daughter cells which function as gametes.

*look at notes for differences in cytokinesis between plants and animals