XBI2M014

UNIVERSITY OF DUBLIN
TRINITY COLLEGE
Health Sciences

BIOCHEMISTRY

Dr. Richard K. Porter

Junior Freshman Medical Year &

2nd Year Radiotherapy students Trinity Term 2010

Wednesday 5 May 9.30 – 12.30 R.D.S.

INSTRUCTIONS:

Answer Question 1 and any other four questions.

(Do not attempt any extra questions.)

ALL QUESTIONS CARRY EQUAL MARKS

N.B.
PLEASE ANSWER EACH QUESTION IN A
SEPARATE ANSWER BOOK.

Items required for this examination: student ID card, pen(s), pencil(s),

eraser, calculator and ruler.

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1. Amphetamine is a reversible inhibitor of monoamine oxidase (the inhibition

shows no time dependence and is reversible on dialysis). Using
radiolabelled serotonin (5-hydroxytryptamine) as a substrate at the
concentrations shown in Table 1, initial rates were measured after a 30
minute incubation. Product formation (5-hydroxyindoleacetaldehyde) was
also measured after 15 minutes incubation of enzyme with substrate to
demonstrate that linear initial rates were measured. Also shown in Table 1
are initial rates at the different concentrations of serotonin in the presence
of various concentrations of amphetamine. Construct a series of double
reciprocal plots (reciprocal initial rate against reciprocal concentration of
serotonin) for 0, 2,4 and 8 mM amphetamine. What type of inhibitor is
amphetamine? (10 marks). Calculate the apparent Km for serotonin and
the apparent Vmax for the reaction (50 marks).

Calculate the inhibitory dissociation constant (Ki) from a replot of the slope
of the double reciprocal plot against the concentration of inhibitor (40
marks).

Table 1. Monoamine oxidase activity as a function of substrate (serotonin)

and inhibitor (amphetamine) concentration
amphetamine concentration (mM)

0 2 4 8
Final concentration of initial rate of mononamine oxidase
serotonin in assay mix (M) activity (nmol/min/mg)
50 33 25 20 14
100 50 40 33 25
200 66 57 50 40
400 80 73 67 57
800 89 84 80 73

2. “In Vitamin B12 deficiency a megaloblastic anaemia identical to

that seen in folate deficiency is observed but, in addition, there is
a neuropathy also associated with vitamin B12 deficiency”.
Discuss the biochemical rationale behind the above statement.

3. Describe the structure of high-density lipoproteins (HDL’s) and

discuss their role in cholesterol homeostasis.

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4. Write an essay on inflammatory response describing the different

processes involved. State why it is important for the medical
profession to understand these processes.

5. Discuss the role of MAP kinase (MAPK) pathways in the

development of cancer.

6. A number of enzymes play a role in DNA synthesis at the

replication fork. Discuss their individual functions in this process.

7. Discuss, with examples, how the initiation of transcription is

regulated in mammalian cells.

8. Discuss the mechanism by which adrenalin stimulates the

production of intracellular 3',5'-cyclic AMP (cAMP).

©University of Dublin 2010