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Tell them what you’re going to tell them. Tell them.

Tell them what you just told them. ~ Aristotle

1. Explain the process of neurotransmission at the neuromuscular junction.


a. A disease associated with the neuromuscular junction
b. One drug specifically targeting this disease.
c. The mode of action of this drug.
Intro Neuromuscular junction is the site where neurons excite skeletal muscle fibres. It is the point of contact of
axon terminals (pre-synaptic terminal) of motor neurons and motor end plate (post-synaptic membrane) of
skeletal muscle fibres. Neurotransmission is the process by which signals are sent across the neuromuscular
junction, from the neuron to the muscle fibre, and we are now going to take a look at how this happens, a
disease that impacts it, and pharmaceutical treatments of this disease
Body Neurotransmission occurs through a sequence of inter-related steps.
- It starts with an action potential travelling the length of the axon of a motor neuron to an axon terminal.
- As the action potential reaches the terminal bouton, it activates voltage-gated calcium channels thus
allowing calcium ions to diffuse into the bouton.
- Calcium ion entry then triggers synaptic vesicles to release acetylcholine via exocytosis. As the vesicles reach
the membrane of the neuron, they release ACh into the synaptic cleft.
- ACh diffuses across synaptic cleft and binds to acetylcholine nicotinic receptors which contains ligand- gated
cation channels on the post synaptic membrane.
- As the ACh binds to the receptors, ligand-gated cations channels open to allow the flow of sodium ions into
the muscle fibres, with some potassium ions flowing out from the muscle fibres. Greater influx of Na+ relative
to efflux of K+ causes membrane potential (-80mV) to become less negative thus depolarizing the motor end
plate & producing end- plate potential. Skeletal muscle cell = -95mV, smooth muscle cell = -80mV neurons = -
70mV
- Once membrane potential reaches a threshold value, an action potential propagates along the sarcolemma.
The voltage of the action potential open voltage gated sodium channels, allowing even further flow of
Sodium into the muscle fibre and propagating the action potential along the fibre. The fibre has now been
stimulated.
•Local current flow between depolarized end plate and adjacent muscle cell membrane brings adjacent areas
to threshold
•Action potential is initiated and propagated throughout muscle fibre

In order to cease neural transmission and reset the synaptic cleft, acetylcholinesterase (AChE) an enzyme
located in the synaptic cleft, destroys the ACh molecules, enabling the closing of the nicotinic receptor bond
cation channels, terminating the muscle response. Choline is transported into the axon terminal for
resynthesis of acetylcholine.
However there are circumstances where these steps do not always flow smoothly. One example of this is
Myasthenia Gravis, an autoimmune disease whereby the body produces antibodies that blocks these
Nicotinic Receptor Cation channels normally opened by the ACh. The result is that ACh has limited receptors,
and therefore limited success at stimulating the muscle. The overall result for the patient is extreme muscle
weakness and fatigue.
But the condition is not without treatment. One of the most common pharmaceutical treatments of
Myasthenia gravis is AChE Inhibitors ie. Neostigmine. By inhibiting the AChE in the synaptic cleft, it allows ACh
longer time to stimulate the few receptor channels that are available to it. This in turn allows for increased
muscle stimulation, closer to normal levels.
However AChE inhibitors are not without their side effect and can cause issues in the GIT like hypermotility.
This could lead to the need to take even further medication to counteract the side effects.
If the effects of the AChE inhibitor begin to lessen, or if the side effects begin to increase too much, then in
general the next option is plasmapheresis, whereby the antibodies causing the issue are extracted from the
blood completely. With reduced antibody levels, the symptoms of Myasthenia Gravis are often reduced,
however it can become a repetitive process.
Conclusion So as outlined, the neurotransmission across the neuromuscular junction is a well-choreographed mutli-step
process. However with conditions such as Myasthenia Gravis, there can be issues with the process, and while
symptomatic treatment is available, there is no cure yet.
Tell them what you’re going to tell them. Tell them. Tell them what you just told them. ~ Aristotle

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Tell them what you’re going to tell them. Tell them. Tell them what you just told them. ~ Aristotle

2. Describe the characteristics of the biomolecules of the cell membrane.


a. Explain the ‘fluid mosaic model’ of the cell membrane.
b. Give an example a membrane transporter.
Intro Cell membrane is the barrier to entry into the cell & maintains cell’s integrity. The main biomolecules of cell
membrane are phospholipids, cholesterol, membrane proteins and membrane carbohydrates, each with
their own characteristics and functions. Over the next few minutes we’ll take a look at some of these
biomolecules, and how they interact.
Body The most abundant biomolecule in the cell membrane are the phospholipids, they constitute a large area in
the cell membrane. Phospholipids are amphipathic meaning that they possess both hydrophobic and
hydrophilic characteristics. This is ideal for its function as barrier for the cell, with the hydrophilic phosphate
heads facing outwards, interacting with the cytoplasm & exterior of the cell while hydrophobic tails point
inwards, interacting with each other generating the impermeable lipid bilayer.
Next is cholesterol, which is also a type of lipid. Cholesterol enhances both the structural integrity of the cell
membrane, as well as aiding in maintaining its fluidity. Cholesterol, like the phospholipids, has a hydrophilic
head and a hydrophobic tail, but in between it has a rigid steroid ring structure (3 6-membered ring, 1 5-
membered ring) with is the bases with its structural characteristics. These cholesterol molecules sit in the cell
membrane between adjacent phospholipids.
Apart from lipid, proteins are also part of the cell membrane. There are two main types of membrane
protein, integral (pass through the plasma bilayer) and peripheral protein that sits in either one side of the
membrane or the other. The membrane proteins carry out many roles and functions including anchoring and
identification of the cell, gated and leak channels, and as enzymes and receptors. The membrane proteins aid
the cells interaction with other cells and with its own surrounding.
The last biomolecule that we will talk about are the membrane carbohydrates. Membrane carbohydrates
exist in many forms, including as part of complex molecules such as glycolipids and glycoproteins depending
on its attachment either to protein or lipids. such as glycocalyx, which is involved in the lubrication,
protection and locomotion of cells, amongst other functions.
The combination of these biomolecules creates the plasma membrane surrounding all cells. The arrangement
of the molecules gives the membrane a 2-dimensional mosaic appearance. But the biomolecules are in some
way restricted in their movement within the membrane. Phospholipids for example may move alone their
over layer with relative ease, but are resisted from rotated within the membrane to point inwards, or vice
versa. This concept of being a changeable 2D mosaic layer both pointing inwards and point outwards is
referred to as the Fluid Mosaic Model.
An example of a membrane transporter, present in most cell membranes is the Sodium Potassium pump. The
pump is powered by the phosphorylation of the pump enzyme by 1 molecule of ATP, and in the process
enables 3 sodium ions to leave the cells, and 2 potassium ions to enter. Following the exchange the enzyme
become unphosphorylated again, and the channel is free to repeat the process again.
Conclusion The fluid mosaic model as described provides for the protection and function of the cell, as well as its
interactions with neighbouring cells and the world around it, with each of the biomolecules having a part to
play.
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Plus potentially some inclusion of the Membrane Proteins.


Tell them what you’re going to tell them. Tell them. Tell them what you just told them. ~ Aristotle

3. Describe how energy is released from glucose.


a. Explain glycolysis, the TCA cycle and ETC.
b. Compare and contrast between the energy released under anaerobic and aerobic conditions.
Intro Mitochondria is the powerhouse of the cell. This expression is more than just a meme, it describes the
function of mitochondria; an organelle found in eukaryotic cells as the site of production of energy for the
body. Over the next few minutes we will see the three processes that convert glucose into energy-carrying
molecules, ATP.
Body The first step is glycolysis. Glycolysis is the metabolic pathway that converts glucose to pyruvate. Pyruvate is
the metabolic precursor of Acetyl CoA that enters the TCA cycle in aerobic respiration. As part of the
conversion, the free energy released is used to form the high-energy compounds ATP and NADH. The
pathway is a sequence of 10 enzyme-catalysed reactions. The intermediates that exist between steps, also
allow for other monosaccharaides such as fructose and galactose to enter the process, as they can be
converted separately to one of the intermediates. The pathway can be split into two phases, the preparatory
or investment phase in which ATP is consumed, and the pay-off phase in which ATP is produced. Since
glucose is a hexose (6 carbon sugar), it’ll lead to 2 triose sugars being produced per glucose molecule. This
duplication leads to a net gain of 2 NADH and 2 ATP molecules per glucose molecule. The process can take
place either aerobically or anaerobically and this process takes place in the cytosol of the cell, not yet in
mitochondria.
The Tri-carboxylic Acid Cycle, or Krebs Cycle, is a series of chemical reactions used by all aerobic organisms to
generate energy through the oxidation of acetate derived from carbohydrates, fats and proteins into carbon
dioxide and chemical energy in the form of ATP. Following on from the Glycolysis of Glucose, in the presence
of oxygen, the 2 pyruvate molecules are oxidised to acetyl CoA in mitochondrial matrix, and reduce a
molecule of NAD+ into NADH. These products are then fed into the Krebs Cycle. As part of the cycle, citric
acid (a type of tricarboxylic acid) is consumed and then regenerated by this sequence of reactions to
complete the cycle. Along the cycle, NAD+ is reduced into NADH, ADP is turned into ATP (1 molecule per
pyruvate), FAD is reduced into FADH2 and as with the glycolysis cycle, as there are 2 molecules of pyruvate,
the cycle occurs twice per glucose molecule.
The resulting molecules of the TCA cycle (10NADH + 2FADH2) go on into the Electron Transfer Chain (ETC)
embedded in inner mitochondrial membrane to undergo oxidative phosphorylation. In the ETC each NADH
will be indirectly responsible for the production of 3ATP and the FADH2 will be responsible for the production
of 2ATP. The net result of the which is 10NADH x 3 = 30 ATP, & 2FADH2 x 2 = 4ATP, with a total of 34ATP. As
we already had 2ATP from the glycolysis cycle, and a further 2 from the Kreb cycle, the total output is 34 + 2 +
2 = 38 ATP. However this is all in the presence of oxygen, i.e. Aerobic reactions.
In the absence of oxygen ie anaerobic reactions, the situation is quite different in the sense that pyruvate
oxidation in the mitochondrial matrix cannot take place and so the pyruvate is converted to lactic acid. We
know that glycolysis produces pyruvate and 8ATP molecules but in the production of lactic acid, 6 ATP
molecules are consumed, making the net production of ATP a mere 2ATP, a considerable difference from this
Aerobic counterpart.
Conclusion For a detailed understanding of each of the steps involved one would need to spend more than 5 minutes,
but the brief outline just given will hopefully shed some light on the basic overarching steps involved in the
aerobic and anaerobic production of energy in the body.
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Aerobic
Reaction Path Intermediate ATP Produced
Glycolysis 2 ATP 2
2 NADH + H+ 6
Pyruvate -> Acetyl CoA 2 NADH + H+ 6
TCA Cycle 6 NADH + H+ 18
2 FADH2 4
2 ATP 2
Total 38

Anaerobic
ATP Produced
Glycolysis 2 ATP 2
2 NADH + H+ 6
Pyruvate -> Lactic Acid -6
2
Tell them what you’re going to tell them. Tell them. Tell them what you just told them. ~ Aristotle

4. Describe the length-tension relationship for skeletal muscle


a. Explain the role that this relationship plays in the heart
Intro The contraction of muscles can be described based on two variables; length and force. Force can be divided
into either tension (force exerted by the muscle on the object) or load (force exerted by the object on the
muscle). When we discuss muscle contraction, we generally refer to two types of contraction which are
isometric contraction (when tension is developed but the overall length of the muscle does not change/
whereby the length of the muscle does not change) and isotonic contraction (where tension increases until it
equals the weight to be lifted then shortens and tension stays constant/ whereby the tension in the muscle
does not change)
In natural movements, the body create multifaceted contractions varying of time between isotonic and
isometric, in order to produce the ideal length and tension relationship. We will now look at this relationship
for skeletal muscle and for cardiac muscle.
Body In muscles, the sliding filament theory from Huxley model outlines how myosin and actin filaments interact to
produce the contraction of the muscle. However, myosin and actin filaments have finite length (sarcomere
length) and so, the amount of overlapping between the filaments would also be finite and could impact their
interactions.
In a relaxed muscle fibre, actin filament has pulled all the way out to the end of myosin filament therefore no
actin –myosin overlap. This cause the tension developed by activated muscle to be zero.
As the sarcomere begins to shorten, actin filament begins to overlap with myosin filaments thus increasing
tension progressively. Further shortening of sarcomere allows all actin filament to overlap with myosin
filament and sarcomere maintains full tension. However, in an extremely shortened muscle, where 2 Z disks
of sarcomere touch the ends of myosin filaments, 2 thin actin filaments begin overlapping each other in
addition to overlapping with myosin filaments thus producing minimal contraction.
This length-tension relationship can be graphed to show the ideal length, which lies close to the natural
resting length of most muscles, as our bodies have evolved for this efficiency.
As the muscle is stretched, the elastic properties of the muscle itself, its connective tissue, ligaments and
tendons, as well as elements such as the titin filaments (located between myosin tick filament and the Z disk)
begin to create a second tension, a passive tension, unrelated to the stimulated/active tension created in the
muscle. This passive tension increases as the muscle is stretched further, as a form of resistance to the
stretching. The sum of the passive tension and the active tension can be graphed together as the total
tension developed while stretching the muscle fibre.
The length-tension relationship can also be seen in cardiac muscle, however with some slight differences. The
resting length of the cardiac muscle occurs after the heart has contracted, but before it starts to fill again. The
filling of the ventricles causes stretching of the cardiac muscle beyond its resting length. While in skeletal
muscle, we would start to see a reduction in the tension produced at this increased length, in cardiac muscle
there is actually an increase in tension. As with skeletal muscle, there is a limit to this increase. For cardiac
muscle the limit comes at approximately the point when the ventricles are completely full (end of diastole),
and just prior to contraction of the ventricles in the cardiac cycle.
The result of this is that there is a higher level of tension created in cardiac muscles when they are slightly
stretched by the preload filling of the heart, then they would be at their natural resting length. It also creates
a more efficient contraction of the ventricles, and the natural stretching of the cardiac muscle, in enable
increased tension creation, and therefore more forceful contraction of the ventricle, aiding circulation, and
helping to push back against vascular resistance.
Conclusion So while the length tension relationship can be observed in both skeletal and cardiac muscle, the ideal length
for skeletal muscle lies near its resting length, while the ideal length for cardiac muscle, lies closer to the
stretched length created by the filling of the ventricles.
Tell them what you’re going to tell them. Tell them. Tell them what you just told them. ~ Aristotle

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Tell them what you’re going to tell them. Tell them. Tell them what you just told them. ~ Aristotle

5. Discuss the critical role fluid balance plays in our bodies by considering the following headings:
a. Describe how body water is compartmentalized in the body
b. Describe the ionic composition of the body’s fluids
c. Explain what osmotic pressure means.
d. What determines the osmotic pressure of body fluids?
e. What unit is used to measure osmotic pressure?
f. Explain why changes in body fluid balance are so dangerous?
g. Detail what happens in the body as a result of dehydration of the body’s fluids.
Intro Approximately 60% of our body weight is fluid, and ensuring a balance of fluid in our bodies is critical and
over the next few minutes we are going see how our body regulated and balances the fluid levels.

Fluid constitutes 60% of our body weight and in newborn the percentage is even higher (75%). Therefore the
constancy of fluid composition is crucial for cell function. Over the next few minutes, we are going to see how
our body regulates and balances fluid levels.
Body Of the fluid, 2/3rd is intracellular, and 1/3rd is extracellular (predominantly in the form of interstitial fluid) with
just 20% of ECF as Plasma. Apart from water, these fluids consist mainly of electrolytes, which are almost
completely disassociated, and some negligible quantities of other biomolecules like amino acids and glucose.
In the ECF the electrolytes comprise of mainly sodium and chloride ions, while in the ICF the electrolytes are
comprised mainly of potassium and phosphate ions as well as proteins that are negatively charged. ECF
composition is critical for cell function, and is maintained by homeostasis, while ICF composition has
important influences on cellular reactions (bcs ionic strength has an important influence on cellular reaction)
and is maintained by mechanism within the cell itself.
Osmotic pressure of body fluids is a measure of the tendency for water to move into solution based on its
relative concentration of non –penetrating solutes and water. The higher the osmotic pressure of a solution,
the lower the water concentration and the higher the solute concentration of the solution. The electrolyte
concentrations in ECF and ICF play a large role in their osmotic pressures, in particular sodium and chloride
ions in (ECF) and potassium and phosphate ions in (ICF). The osmotic pressure of ECF and ICF is roughly equal,
resulting in no net movement of water between the two. Thus, ensuring that cell volume remains constant.
The osmotic pressure exerted by particles in a solution is determined by the number of the particles per unit
volume of fluid not by the mass of the particles and the unit used is osmole. Osmole refers to number of
osmotically active particles in a solution rather than molar concentration. 1 osmole (osm) is equal to 1 mole
(mol) of solute particles. However, the osmole is too large a unit to express osmotic activity of solutes in body
fluids, so milliosmole is used (mOsm). 1x10^-3

The exact osmolarity of body fluids is 283mOsm/L and control of free water balance is critical for regulating
body fluid osmolarity. Any changes to body fluid osmolarity can result in movement of water into or out of
the cells as water moves from a solution of low osmolarity to one of higher osmolarity until osmolarity in the
two solutions are equal. This can disrupt cellular function, and put additional strain on the body’s organs.
In cases of dehydration, water content of plasma and interstitial fluid (the entire ECF) decreases causing
increased concentration of ECF with a related increase in osmolarity of ECF. Water will then leave all cells by
osmosis through the cell membranes in order to increase osmolarity of ICF. This can cause disruption of
cellular function. Symptoms are mainly neurological as water loss from brain cells leads to shrinkage of cells.
Conclusion So we have seen how important it is to maintain the body fluid balance to avoid any problems that could
arise in the failure to do so, worst could be circulatory shock and death.
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6. Explain the processes of transcription and translation.


Intro DNA or Deoxyribose Nucleic Acid is a molecule that holds the majority of the genetic information required for
the development, operation and reproduction of all living organisms. DNA is copied over and over again
through a process known as Replication, most notably during the S Phase of the cell cycle, prior to cell
division. But replication is not the only process that DNA undergoes, and we are now going to look at two of
the other processes, transcription and translation.

DNA, Deoxyribose Nucleic Acid is a genetic material. Our genetic information is encoded by the sequence of
different nucleotide bases in DNA and is responsible for the development, operation and reproduction of all
living organisms. DNA synthesis occurs by the process of replication that happens most notably during the S
phase of cell cycle. But in order to produce functioning molecules for the body to function, DNA has to
undergo two more processes; transcription and translation.
Body Transcription is the process where Ribose Nucleic Acid, or RNA are synthesised from a DNA template and it
happens in 5’ to 3’ direction of mRNA (transcription occurs in 3’ to 5’ direction on DNA template –antisense
strand). Transcription occurs in 3 steps.
Steps of transcription:
The first step is initiation. An enzyme, RNA Polymerase, in closed complex form, binds to a promoter region
on the DNA which identifies the start of the DNA sequence to be transcribed. The RNA Polymerase then
changes to Open Complex form and in doing so unwinds a 17-18 base pairs segment of the DNA.
Next is elongation: RNA Polymerase moves along the “antisense” strand of the unwind DNA template
synthesizing mRNA until it reaches the terminator region. RNA Polymerase reads antisense strand of DNA in
3’ to 5’, making mRNA copy 5’ to 3’, matching the DNA bases with their RNA counterparts, similar to the
matching of base pairs in DNA Replication, but with the notable difference that DNA’s thymine is replaced by
uracil in RNA.
Lastly is termination: When the RNA Polymerase reaches the Terminator region of the DNA strand, it causes
the RNA Polymerase to stop, disassociate from the DNA and release the mRNA. Forces in the DNA cause it to
wind itself back together again, returning it to its original form.
The mRNA however is not the end product; it will then undergo translation. Translation is the synthesis of
proteins directed by a mRNA template in a ribosome with the help of tRNA. The Ribosome itself is made up of
Ribosomal RNA and a variety of proteins, and is split into 2 units, the small unit and the large unit.
Translation also occurs in 3 steps;
Initiation: Amino acids are matched to the mRNA through in sets of 3 nucleobases known as codons. The
small and large ribosomal units assemble around the mRNA. Charged initiator tRNA that is tRNAMet enters
the P site and binds to the start codon AUG which code for methionine on the large subunit. AUG is the
codon for amino acid Methionine or the start codon. The variable portion of the protein starts after that. The
protein chain begins.
Elongation occurs by the binding of the next aminoacyl-tRNA to the large unit of ribosome in line with each
next codon in turn. In this way ribosome is repeatedly moving in steps of 3 nucleobases at a time along the
mRNA. Peptidyltransferase reaction takes place, forming bond between the previous amino acid & incoming
amino acid and the peptide chain beings to grow. As each amino acid joins to the chain the tRNA linked to
the previous amino acid detaches and uncharged tRNA exits the ribosome through E site.
Termination: The peptide chain growth continues until the ribosome reaches a “Stop Codon” (UAA, UAG,
UGA). The tRNA linked to the stop codon contains no amino acid, and therefore when the penultimate tRNA
detaches from its associated amino acid, the peptide chain is released from the P site of ribosome and the
ribosome dissembles from the mRNA.
As each codon is made up of 3 nucleobases, and there are 4 different RNA nucleobases, this allows for 64
different codons (4x4x4). However, there are only 20 standard amino acids, meaning that some amino acids
link to more than 1 Codon. But if you were to take a section of mRNA that codes for 10 codons, the first being
AUG/Methionine and the last being the Stop codon, despite its small size, it can still code for in excess of 25
billion different amino acid combinations.
Conclusion Therefore, while there are several intricate steps to be taken to convert a DNA sequence into a protein, the
end results and possible combinations are vast. However the number and intricacies of the steps required
also allow for several sites for mutations to occur and this may cause deadly effects.
Tell them what you’re going to tell them. Tell them. Tell them what you just told them. ~ Aristotle
Translation- Elongation
Transcription
Images to Promoter Transcription start site Ploy A termination
Transcription start site Stop codon
Draw 3’ 5’
5’ 3’ 5’ 3’ mRNA
Transcription factors bind A U G U U UG G AG U G … U G A
TF A A A C C U
3’ 5’
5’ 3’
RNA polymerase
Transcription elongation
3’ 5’
5’ 3’
RNA polymerase
Met Phe Gly

3’ 5’
DNA
5’ 3’
5’ 3’ mRNA Translocation
Ribosome moves along 3 bases and mRNA
specifies next amino acid (aa). tRNA
RNA sequence is complementary to template strand
Tell them what you’re going to tell them. Tell them. Tell them what you just told them. ~ Aristotle

7. What are enzymes and how do they work?


a. Describe what happens to the velocity of a reaction as you increase the substrate concentration.
b. Discuss other factors that can affect enzyme function.
c. Explain what is meant by competitive and non-competitive inhibitors using graphs to illustrate this.
d. Describe what the Michaelis constant is, and how this relates to affinity.
Intro The human body relies heavily on the highly selective catalysts called enzymes in order to perform chemical
reactions and function. Enzymes are biological catalyst that accelerates a chemical reaction in the cells
without being consumed in the process. They do this by creating an alternative reaction pathway with lower
free energy of activation. Enzymes do not change the equilibrium of a reaction or its energy balance, they
only make the reaction go faster. We will now take a brief look at some of the factors involved in reaction
with enzymes.
Body Simple reactions often show linear relationships between the rate of product formation, and the
concentration of the substrate, so as the concentration substrate increase there is a linear increase in the
rate of product formation. But in enzyme-catalysed reactions this is not the case. Mot enzymes have a
hyperbolic kinetic curve to show the relationship between velocity against substrate concentration.
At low substrate concentrations, there is an abundance of enzyme, and the speed of the reaction is limited by
the amount of substrate available. But as the substrate increases, the speed of the reaction increases quickly,
and greater speed than without the enzyme.
Rate increases with concentration of substrate until a maximum velocity is reached, known as Vmax where
then all active site of the enzymes are saturated and speed of reaction plateaus and is limited by the speed of
enzyme function itself.
The amount of substrate concentration required to reach ½ of Vmax is known as the Michaelis Constant and
is denoted by Km.
Apart from substrate concentration, temperature and pH can also affect enzymatic function due to
denaturation of the enzyme at non-ideal temperature or pH levels. Reaction velocity and enzyme activity will
increase, up to a peak point of both velocity/temperature curves and velocity/pH curves. Beyond those
respective points, denaturation of the enzyme causes the rate of reaction to actually decrease, hence it is
best to maintain the reaction within certain temperature and pH limits to ensure optimal speed is achieved.
In addition to Temperature and pH, the presence of inhibitors can also affect the rate of the reaction. There
are two types of inhibitors; competitive and non-competitive. Competitive inhibitors binds to the same site as
the substrate that is the active site thus inhibits the enzyme from catalysing the substrate reaction. This
however can be overcome by increasing the substrate concentration, and essentially forcing the competitive
inhibitor out of the way, eventually allowing the substrate access to the enzyme again. The net effect of this
is an increased Km because more substrate is needed to reach ½ of maximum velocity, but Vmax remains the
same as the enzyme still becomes saturated by the substrate at high concentration levels
Non-competitive inhibitors on the other hand do not bind to the same site on the enzyme as the substrate.
They instead bind to an alternative site; the allosteric site and the inhibitor changes the conformation of the
active site also resulting in the inability of the substrate to bind to active site of enzyme. This kind of
inhibition cannot be overcome by the substrate, even at high concentration levels because these inhibitors
essentially reduces the amount of functioning enzyme available to catalyse the substrate reaction.
The net effect of this is a decreased Vmax due to less active enzyme molecules. The reactivity of the substrate
and the enzyme however is unchanged, and therefore they remain with the same Km.
Comparing the Michaelis Constant, Km, of two enzymes of the same substrate allows you to compare the
affinities of the 2 enzymes for the substrate. The Michaelis Constant has an inverse relationship with affinity,
in that the lower the Km, the higher the affinity, and the higher the Km, the lower the affinity. This can be
explained as a higher Km, is the result of requiring a higher concentration of substrate in order to get the
enzyme to react quickly, i.e. requiring to flood the enzyme before it reacts. An enzyme with greater affinity, is
more willing to react and therefore requires lower concentrations of the substrate in order to react quickly,
and therefore has a lower Km.
Conclusion So as outlined, enzymes are proteins that help to catalyse reactions in the body and the enzymatic function
are highly dependent upon temperate and pH of the reaction, the substrate concentration, presence of
inhibitor and the affinity of the enzyme to the substrate.
Tell them what you’re going to tell them. Tell them. Tell them what you just told them. ~ Aristotle

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Tell them what you’re going to tell them. Tell them. Tell them what you just told them. ~ Aristotle

8. Receptor signalling and ligand-receptor interactions. Using tamoxifen as an example, discuss receptor signalling and
inhibition. Tamoxifen is a partial agonist on the oestrogen receptor and is used in the treatment of breast cancer.
a. What are an agonist and an antagonist?
b. Describe the structure of the oestrogen receptor?
c. Describe the events that follow when oestrogen binds to its receptor?
d. What is a partial agonist and what is the consequence of using a partial agonist such as tamoxifen?
e. Is there an option of using a pure antagonist?

Intro As the name implies, receptors are protein molecules that receive chemical signals from outside a cell.
Receptor binding triggers intracellular signalling cascades with protein –protein interactions and enzymatic
reactions. These cascades regulate metabolic enzymes, membrane transporters, ion channels and genes. We
will now be looking at a specific case of this, in the oestrogen receptor, which is one of the receptors targeted
in the treatment of breast cancer. Without treatment, the receptor, on binding with oestrogen, encourages
breast cancer cells to multiple.
Body First, we will look at the stimulation of these receptors. Hormones or other extracellular signalling molecule
that binds to receptors are said to be either agonists or antagonists. In their simplest form agonist stimulate
the receptors, causing biological response while antagonists inhibit the stimulation of receptors. Agonists
acting as accelerators, antagonists acting as brakes. They can work together acting on different receptors or
in competition for the same receptors.
Oestrogen is a type of lipophilic hormone, it uses intracellular gene –specific transcription factors. The
intracellular receptors for this hormone is structurally similar and are referred to as the steroid hormones
superfamily of receptors and are primarily localised to the nucleus thus the oestrogen receptor is called a
nuclear receptor, which comes in 2 forms, alpha and beta. Both forms contain the same 5 sections along its
length as follows:
- The N Terminal domain, including Activation Function 1
- The DNA Binding Domain (DBD)
- The Hinge domain
- The Ligand Binding Domain, including Activation Function 2
- And the C Terminal Domain

After the oestrogen binds to its receptor, the ligand binding domain changes its shape and displaces the heat-
shock protein (chaperon protein) complex that had been inhibiting its action. With the protein complex
displaced it allows for the binding of cofactors (both co-activators and co-repressors) to the receptor, which
help to promote its interaction with DNA target genes, making use of both Activation Function 1 and 2 (AF1
and AF2). Oestrogen binds to the receptor like a lock and key after it has passively diffused into the cell
forming a hormone receptor complex then moves into the cell nucleus through nuclear pores.
Partial agonists are drugs that bind to and activate a given receptor, but have only partial efficacy at the
receptor relative to a full agonist. They may also be considered to display both agonistic and antagonistic
effects - when both a full agonist and partial agonist are present, the partial agonist actually acts as a
competitive antagonist, competing with the full agonist for receptor occupancy and producing a net decrease
in the receptor activation observed with the full agonist alone. In the case of Tamoxifen, the natural
oestrogen in estradiol when bound the oestrogen receptor promotes a fully activated transcription, both AF1
and AF2, whereas Tamoxifen only promotes a partially activated transcription (via AF1). Meaning that there is
still some DNA transcription taking place in the breast cells thus it can be useful to treat early and advanced
oestrogen recptor- positive (ER+) breast cancer because breast cancer cells require oestrogen to grow.
As an alternative, Fulvestrant is a pure antagonist used in treatment of advanced breast cancer in
postmenopausal women. It blocks dimerization and nuclear localisation of the oestrogen receptor, reduces
cellular levels of oestrogen receptor and blocks gene transcription completely (neither AF1 nor AF2)
-have similar structure to oestrogen, can interact with immunoassays fro blood estradiol concentrations and
show falsely elevated results thus can improperly lead to discontinuing the treatment.
Conclusion While Tamoxifen goes some way to inhibiting the action of Oestrogen at the Oestrogen Receptors in breast
cancer cells, it is not without its issues, firstly that it is only a partial agonist, leads room for further issues, but
in addition to that, it also has been found to have differing effect on different tissue types, where in the
Endometrium it has been found to actually increase the risk of uterine cancer.
While Fulvestrant improves on some of these issues, it has other issues itself, one primary example being the
Tell them what you’re going to tell them. Tell them. Tell them what you just told them. ~ Aristotle

cost, which can be several hundred times greater than alternative solutions. These elements would have to
be considered when taking into accounts a patient ability to fund their treatment.
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SERMs- selective estrogen


receptor modulators
Tell them what you’re going to tell them. Tell them. Tell them what you just told them. ~ Aristotle

9. Compare and contrast Ipratropium and Salbutamol in the treatment of patients with bronchoconstriction.
a. Use your knowledge of agonists and antagonists to explain how these drugs serve to alter sympathetic tone
in the lungs.
Intro Bronchoconstriction is found to occur in conjunction with several medical conditions, one of the more
common and widely known conditions being asthma. In bronchoconstriction, the lung airways constrict due
to the tightening of surrounding smooth muscle. Loads of factors could cause the tightening of the muscles
but inhibiting the muscle tightening is one of the primary treatments in reversing bronchoconstriction. We
will now look at two of the pharmaceutical treatments used in inhibiting or reversing bronchoconstriction,
Salbutamol and Ipratropium.
Body Salbutamol is referred to as a beta 2 agonist. To start off, we need to look at the different types of receptors
in the body. The two main categories are Nicotinic Receptors and Muscarinic Receptors, both are adrenergic
receptors. (adrenoreceptors are G-protein coupled receptors) The Nicotinic receptors are further subdivided
into Alpha and Beta-receptors, and both of these, as well as Muscarinic receptors, and further subdivided to
5 different types. These receptors are found in varying levels through the body, and are used in the control of
varying different bodily functions. Salbutamol has a specific focus on the Beta 2 receptors as its target.
Next, we will look at the stimulation of these receptors. Chemicals and molecules that has effects on these
receptors are said to be either agonists or antagonists. In their simplest form Agonist stimulate the receptors,
acting as accelerators and antagonists inhibit the stimulation of receptors, acting as brakes. They can work
together or in competition for the same receptors, or together acting on different receptors. Salbutamol, as
mentioned, is a Beta-2 Agonist, meaning that it acts to stimulate the Beta 2 receptors. Stimulation of these
receptors causes the muscles in the airway to relax and dilate, causing bronchodilation which is the opposite
of bronchoconstriction
The second pharmaceutical treatment we will look at is Ipratropium, which is referred to as a muscarinic
antagonist. As outlined earlier, Muscarinic receptors is one of the main groupings of receptors, which is
subdivided into M1 to M5. However, unlike Salbutamol, which specifically targets Beta 2 receptor,
Ipratropium targets the entire Muscarinic range, and has been found to do so equally from M1 to M5.
As mentioned Salbutamol acts in a stimulatory capacity as an agonist, whereas Ipratropium acts as an
antagonist, meaning that Ipratropium blocks the stimulation of the muscarinic receptors from naturally
occurring stimulations. The stimulation of the muscarinic receptors in the lungs causes constriction of the
smooth muscles, and as such inhibiting this stimulation will lessen the constriction of the muscles, and the
overall bronchoconstriction.
Conclusion So although Salbutamol, a Beta-2 agonist, and Ipratropium, a muscarinic antagonist target different receptors
in the lungs, and act on the receptors in different ways, they are both used in the inhibition or reversal of
bronchoconstriction. A benefit of their differing modes of action is that they can both be administered
without impeding on the action of the other. It is for this reason that they are at times administered together
in order to increase the chances of reversing the bronchoconstriction.
A drawback is that they also have some similar side effects, including headaches and dry mouth, so a
combined treatment with both, is increasing the likelihood of these side effects. However the benefits, both
of individual treatment, and combined treatment are generally seen to outweigh the drawbacks. So much so
that both medications are included in the World Health Organisation list of Essential Medicines
Images to N/A – Potentially a table just outlining targets or each, and mode of action
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Tell them what you’re going to tell them. Tell them. Tell them what you just told them. ~ Aristotle

10. Discuss the basic biomolecules, proteins, lipids and carbohydrates.


a. Outline the basic building blocks, macromolecular structures and how these are formed.
b. For proteins explain what the primary, secondary, tertiary and quaternary structure means.
Intro There are 4 major classes of biomolecules; lipids, carbohydrates, proteins and nucleic acids. Each of them is
built from different building blocks or monomer, has different structures and is formed differently. We will
now look at each of them individually and compare their differences.
Body First, we’ll look at lipids. Lipids composed of long hydrocarbon chains, bonded together with carbon-carbon
bonds both single and double bonds. They hold large amount of energy and are therefore one of the main
energy storage molecules with the cell. Phospholipids, a class of lipids, possess amphipathic properties which
is crucial for its role as a major component of the cell membrane. Cholesterol and other sterol are also types
of lipid, and they also constitute cell membranes. Differing forms of lipids contain differing bond types,
including ester bonds in triglycerides.
Next up is carbohydrates. They provide energy source for cells, and can also have structural roles. They are
made up of monosaccharaides and can either be in a long straight formation, or in a ring structure common
example would be glucose. The monosaccharaides like glucose, fructose and galactose can link together,
through glycosidic bonds to form disaccharides (like sucrose and lactose) and polysaccharides (such as
cellulose and starch).
Proteins account for the largest portion of the biomolecules present in the cell, and have wide variation. They
have both structural and operation roles, in the forms of enzymes and hormones. Proteins are composed of
monomers called amino acids, which are bound together by peptide bonds in a polypeptide chain. There are
20/21 amino acids in humans, of which ours bodies can synthesise 11 (and we call these the non-essential
amino acids), and the rest (the essential ones) we must consume. Amino acids consist of an amino group, a
carboxyl group and hydrogen, all bonded to a central carbon, alongside a variable residue group/R group for
short. It is the R group that gives each amino acid its special properties, and the combination of these amino
acids into polypeptide chains, will give the protein its function and structure.
There are 4 types of protein structures. The primary structure is the simplest, with just the basic bends in the
straight protein chain where amino acids are linked together with peptide bond. The secondary structure
consists of either alpha helixes or beta pleated sheets. A protein may switch from one section to the next
between alpha helix and pleated, and the combination of these changes creates globular protein which is the
tertiary structure. The quaternary structure is where multiple polypeptide chains are being held together
with non-covalent bond and/or interchain disulphide bonds, such as can be seen with the structure of
haemoglobin.
In addition to these 3 main types, there are Nucleic Acids which are responsible for our genetic code. They
exist most readily in DNA and RNA and are formed from subunits called Nucleotides. Each nucleotide is
composed of a nitrogenous base, a 5-carbon sugar and 3 phosphate groups. There are 5 different types of
Nucleic Acids in the cells; they are Adenine, Thymine, Guanine, Cytosine and Uracil. However nucleotides are
responsible for more than just DNA and RNA. For instance, ATP. It is a nucleotide responsible for the energy
within the cell. In DNA the nucleic acids, and their sugar phosphate backbones make use of phosphodiester
bonds and hydrogen bonds.
Conclusion Each of these 4 biomolecules types: lipids, carbohydrates, proteins and Nucleic Acids, as outlined, are very
different from one another, both in how they are built and in how they operate, however each is equally
important in its own right. There are also many points of interaction between them, such as the formation of
protein based on RNAs Nucleic Acid base coding, and as such none of the 4 should ever be overlooked.
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Tell them what you’re going to tell them. Tell them. Tell them what you just told them. ~ Aristotle