You are on page 1of 4

BIOCHEMISTRY LABORATORY: PROBLEM BASED LEARNING EXERCISES

Amino Acids and Proteins


Answers to Questions:
CASE 1
1. Explain the biochemical basis for the patients disease.
Alzheimers disease is a progressive neurodegenerative brain disease wherein the component of
the amyloid plaque that accumulates is amyloid (A), an extracellular peptide. This peptide, when
aggregated in a -pleated sheet configuration, is neurotoxic, and is the central pathogenic event
leading to the cognitive impairment characteristic of the disease.
The A that is deposited in the brain in AD is derived by proteolytic cleavages from the larger
amyloid precursor protein. The proteolytic cleavage is achieved by the action of the enzyme secretase.
The A peptides aggregate, generating the amyloid that is found in the brain parenchyma and around
blood vessels.
2. Explain the theories involved in the development of Alzheimer disease.
a. Formation and accumulation of amyloid plaques (amyloid hypothesis)
Amyloid is a peptide produced by enzymatic cleavage from amyloid precursor protein. This
amyloid , when aggregated in a -pleated sheet configuration, forms amyloid plaques which is
neurotoxic and causes cognitive impairment.
b. Accumulation of neurofibrillary tangles inside neurons (tau hypothesis)
A second biologic factor involved in the development of Alzheimer disease is the accumulation
of neurofibrillary tangles inside neurons. A key component of these tangled fibers is an abnormal
form (hyperphosphorylated and insoluble) of the tau () protein, which, in its healthy version, helps in
the assembly of the microtubular structure. The defective appears to block the actions of its
normal counterpart.
CASE 2
Clinical impression: SCURVY
1. Explain the biochemical basis for the patients disease.
Collagen contains hydroxyproline (hyp) and hydroxylysine (hyl) which are not present in
most other proteins. These residues result from the hydroxylation of some of the proline and lysine
residues after their incorporation into polypeptide chains. These hydroxylation reactions require
molecular oxygen, Fe+2, and the reducing agent vitamin C (ascorbic acid). If there is deficiency in
ascorbic acid, the hydroxylating enzymes (prolyl hydroxylase and lysyl hydroxylase) are unable to
function leading to lack of prolyl and lysyl hydroxylation. Moreover, interchain H-bond formation
is impaired, as is formation of stable triple helix in collagen.
2. Explain the biochemical basis for the patients signs and symptoms.

Scurvy is a disease characterized by sore and spongy gums, loose teeth, fragile blood vessels,
and poor wound healing. Hydroxylation of proline and lysine is important in the synthesis of collagen.
Hydroxyproline is important in stabilizing the triple helical structure of collagen because it
maximizes interchain hydrogen bond formation. A deficiency in vitamin C leads to lack of
hydroxylation of the two amino acids which now leads to impairment of H-bond formation and
unstable triple helix of the collagen. Additionally, collagen fibrils cannot be cross-linked, greatly
decreasing the tensile strength of the assembled fiber. With this, the connective tissue then becomes
defective, resulting to the above signs and symptoms that characterizes scurvy. Fragile capillary (fragile
blood vessels) can cause subcutaneous extravasation of blood, which explains the multiple hemorrhage
present near the distal end of the patients nail.
***In scurvy, which is due to vitamin C deficiency, hydroxylation of proline residues is decreased, and
an unstable form of collagen is produced. Bones, teeth, blood vessels, and other structures rich in collagen
develop abnormally. Bleeding gums and poor wound healing are often observed.
Reference:

Harvey, R. & Ferrier, D. (2011). Lippincotts Illustrated Reviews: Biochemistry 5th edition.
pp. 20-21, 45-47, 377
Koeppen, B.M. and Stanton, B.A. (2010). Berne and Levy Physiology 6th edition. Mosby, Inc.
(Elsevier): Canada
page 38
CASE 3
1. Explain the difference between human insulin and Humalog mix 75/25 as to structure and
pharmacokinetics.
Chemical Structure:
Insulin contains 51 amino acids arranged in 2 polypeptide chains, Chain A and Chain B.
Chain A is made up of 21 amino acids, while Chain B consists of 30 amino acids.
Residues A7 to B7 and A20 to B19 of chains A and B are covalently bonded by disulfide
bridges.

Structure of human proinsulin (C-peptide plus A and B chains) and insulin. Insulin is shown as the
shaded (orange color) peptide chains, A and B. In insulin lispro, the proline and lysine, at B28 and B29
respectively, have been reversed.
Insulin lispro, the first monomeric insulin analog to be marketed, is produced by
recombinant technology wherein two amino acids near the carboxyl terminal of the B
chain have been reversed in position: Proline at position B28 has been moved to B29,
and lysine at position B29 has been moved to B28.
The Humalog mix 75/25 is a mixture of insulin lispro solution, a rapid-acting blood
glucose-lowering agent and insulin lispro protamine suspension, an intermediate-acting
blood glucose-lowering agent.

Pharmacokinetics:

ONSET

PEAK

DURATION

a. Human regular insulin

30 min 1 hour

2 3 hours

4 6 hours

b. 75/25 Humalog

Less than 15 min

1.5 hours

Up to 10 16 hours

(75% Protamine lispro/25%


Lispro)
If the 2 amino acids in insulin lispro are reversed it would not have any effect with its
half-life, its binding to the insulin receptor or its immunogenicity, which are similar to
those of human regular insulin. On the other hand absorption for the Humalog mix is
faster than regular insulin. Insulin lispro begins to exert its effects within 15 minutes of
administration. Human insulin has a slower onset of action thus; it should be injected 30
minutes to an hour before meals. Peak serum concentrations of Humalog 75/25 occur 90
minutes after subcutaneous administration. With subcutaneously administered human
insulin, peak serum concentrations occur within two to three hours.
2. Explain the biochemical basis for the difference in onset of action between insulin and Humalog
mix 25/75.
Reversal of the proline at B28 and the lysine at B29 results in a more rapid dissolution in insulin
lispro to a dimer then to a monomer that is more rapidly absorbed through subcutaneous
injection.
The very low propensity of 75/25 Humalog serves as its advantage in contrast to human insulin,
to self-associate in an antiparallel structure and form dimers. Lispro dissociates into monomers
almost instantaneously after administration through injection. This characteristic causes a more
rapid absorption as compared with regular insulin giving a shorter onset of action.

REFERENCE:
Katzung, B. (2012). Basic & Clinical Pharmacology. 12th ed. New York: McGraw-Hill Medical p. 744
Brunton, L. (2011). Goodman & Gilman's The Pharmacological Basis of Therapeutics 12th ed.
McGraw-Hill Professional. pp. 1250-1251)

CASE 4:
1. What is the most likely diagnosis? Justify your answer.
Based on the description of the pain that shes experiencing, the patient most likely has sickle
cell disease; which is common among the people with African-American descent.
The pain that the patient is experiencing is due to anoxia because the sickle cells block the flow
of blood into the capillaries, thus, there is interruption in the oxygen supply which explains her fatigue
and the paleness of her mucosal membranes. Her condition was triggered by the urinary tract infection
she is currently experiencing (based on the results of her urinalysis, and white blood cell count, frequent
and burning urination)
2. Explain the biochemical basis for the patients condition.
Single amino acid substitution on the hemoglobin chain. A molecule of HbS contains 2 normal
-globin chains and 2 mutant -globin chains, in which glutamate at the sixth position has been replaced
with valine.
*** Hemoglobinopathies result from mutations that produce alterations in the structure of hemoglobin. There
are many described hemoglobinopathies. One common mutation results in sickle cell anemia, in which the
chain of haemoglobin contains a valine rather than a glutamate at position 6 (designated as E6V, using the
single-letter codes for the amino acids. E6V means that the glutamate [E] at position 6 in the amino acid chain
[where the amino terminal is amino acid number 1] has been replaced by a valine [V]). Thus, in the mutant
hemoglobin (HbS), a hydrophobic amino acid replaces an amino acid with a negative charge. This change
allows deoxygenated molecules of HbS to polymerize. Red blood cells that contain large complexes of HbS
molecules can assume a sickle shape. These cells undergo hemolysis, and anemia results. Painful vasoocclusive crises also occur, and end-organ damage may result.
References:
Denise R. Ferrier, Lippincott's Illustrated Reviews Biochemistry, 6th edition, pages 35-37

You might also like