You are on page 1of 3

Year 10 Pre-Diploma Biology

DENATURING
MEMBRANE
PROTEINS
An investigation:
At what temperature do the channel proteins of
a membrane denature?

Year 10 2014-2015 Pre-Diploma Biology

AT WHAT TEMPERATURE DO MEMBRANE


PROTEINS DENATURE?
Beetroot cells contain an intensely red, water-soluble pigment, Betacyanin. This pigment is soluble in
water and, if cell membranes are damaged, the pigment may escape in sufficient quantities to be
detected in the water around the tissue. (This is what happens when you pick up a cut beetroot and
your fingers become stained red.) One effect of heat on cell membranes is to denature the proteins of
the membrane. Then the proteins will no longer be able to control the diffusion outwards of the red
pigment, so the pigment will be found in the surrounding water.
THE GENERAL QUESTION, WHICH DRIVES THIS INVESTIGATION
At which temperature would you expect the proteins in the plasma membrane of beetroot cells to be
denatured by heat? Write this as the title of the investigation.
HYPOTHESIS
You should be able to propose a hypothesis, based upon your knowledge of proteins, to suggest at
which temperature the proteins will be denatured. Write down a suitable hypothesis and justify it.
TESTING THE HYPOTHESIS THE INVESTIGATION
We will carry out an investigation to determine the approximate temperature at which membrane
proteins are denatured, using identically sized sections from beetroot tissue.
Read all these instructions carefully before you start. Then make a FLOW CHART of the
procedure. When I have seen and confirmed the flow-chart, you may start work.
1
2
3
4
5
6
7

8
9
10

11

Drill out 8 identical cylinders of beetroot with a cork borer, to a length which fits easily into the
available test tubes. You will need to make all cylinders the same length.
Set up 4 different water baths at temperatures between 0c and 75c.
Label 4 test tubes A, B, C & D, according to the temperature of the water baths.
Put distilled water in each of the tubes A, B, C & D. The volume of water is not important as
long as it covers the beetroot tissue. (All you are doing is cooking the beetroot cylinders.)
Place 2 cylinders of beetroot in each of the 4 test tubes.
Place one test tube in each of the water baths for 10 minutes, so that the beetroot tissue is
heated to the temperature of the water bath.
After the 10 minutes of heating, the cylinders of beetroot should be removed from tubes A, B,
C & D and the water thrown away.
While the beetroot samples are heating, collect another 4 test tubes and label them A1, B1, C1
& D1.
Place an identical volume of distilled water in each of the test tubes A1, B1, C1 & D1. The
volume of water must be enough to cover the beetroot samples, but this time it is important that
the volume of water is the same in each tube.
Transfer the beetroot cylinders to their corresponding tubes A1, B1, C1 & D1. The cylinders
will now be covered by distilled water. Leave these samples in a test tube track for a fixed
period of 15 minutes at room temperature. (During this time the pigment betacyanin should
diffuse through the damaged membranes of the beetroot cells. The more denatured are the
proteins, the faster will be the rate of diffusion of the betacyanin.)
After 15 minutes carefully take out the beetroot cylinders and throw them away. Keep the
water! The water contains the pigment which will contain any pigment that has leaked out of
the tissue, through the denatured proteins and is what you will assess.

Year 10 2014-2015 Pre-Diploma Biology

OBTAINING RESULTS AND TREATING THEM


The intensity of the colour of the solutions shows how much pigment is present. Make a scale from 1
to 5 which could be used to estimate the intensity of the red pigment. This is a quantitative scale and is
better than simply describing the colour.
Write down the scale you decide to use.
Now use the scale to estimate the intensity of the pigment in the water in each test tube. Make a table
of your results.
You can confirm the accuracy of your scale with very accurate readings from a colorimeter, using a
blue filter (complementary to red). If there is time, use the colorimeter to confirm your scale of results.
Plot the results you obtain on a graph. Make a line graph it will not have many plots, but the graph
should provide a curve from which you can see the approximate temperature at which the escape of
betacyanin suddenly accelerated. At the temperature where the proteins are denatured and can no
longer control diffusion, there will be a greater density of the pigment.
CONCLUSIONS
Comment upon the results and the graph which you obtained. What does the graph show? What
happened? What is the explanation? If the temperature value for the denaturing of the membrane
proteins is different from the one proposed in your hypothesis, comment on the possible explanations
by analysing your experimental method, or reviewing the hypothesis.

Technician: each group probably requires:


Beetroot
Scalpel
Cork borer
Scissors
8 boiling (test) tubes
Forceps
Equipment for water baths, incl. ice & boiling water
Graduated, measuring cylinder
Distilled water
Cutting surface

John Osborne
May 2015

Year 10 2014-2015 Pre-Diploma Biology

You might also like