Year 10 Pre-Diploma Biology

DENATURING MEMBRANE PROTEINS

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

Year 10 2013-2014 Pre-Diploma Biolo !

AT WHAT TEMPERATURE DO MEMBRANE PROTEINS DENATURE?

Beetroot cells contain an intensel! red" water-soluble pi ment" Betacyanin# $his pi ment is soluble in water and" if cell membranes are dama ed" the pi ment ma! escape in sufficient %uantities to be detected in the water around the tissue# &$his is what happens when !ou pic' up a cut beetroot and !our fin ers become stained red#( )ne effect of heat on cell membranes is to denature the proteins of the membrane# $hen the proteins will no lon er be able to control the diffusion outwards of the red pi ment" so the pi ment will be found in the surroundin water# THE GENERAL QUESTION, WHICH DRIVES THIS INVESTIGATION At which temperature would !ou e*pect the proteins in the plasma membrane of beetroot cells to be denatured b! heat? Write this as the title of the investigation. HYPOTHESIS You should be able to propose a h!pothesis" based upon !our 'nowled e of proteins" to su est at which temperature the proteins will be denatured# Write down a suitable hypothesis and justify it. TESTING THE HYPOTHESIS THE INVESTIGATION +e will carr! out an in,esti ation to determine the appro*imate temperature at which membrane proteins are denatured" usin identicall! si-ed sections from beetroot tissue# .ead all these instructions carefull! before !ou start# Then make a FLOW !"#T of the procedure# +hen / ha,e seen and confirmed the flow-chart" !ou ma! start wor'# 1 2 3 4 4 : 3 Drill out 0 identical c!linders of beetroot with a cor' borer" to a len th which fits easil! into the a,ailable test tubes# You will need to ma'e all c!linders the same len th# 1et up 4 different water baths at temperatures between 02c and 342c# 5abel 4 test tubes A" B" 6 7 D" accordin to the temperature of the water baths# Put distilled water in each of the tubes A" B" 6 7 D# $he ,olume of water is not important as lon as it co,ers the beetroot tissue# &All !ou are doin is 8coo'in 9 the beetroot c!linders#( Place 2 c!linders 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 heatin " the c!linders of beetroot should be remo,ed from tubes A" B" 6 7 D and the water thrown awa!# +hile the beetroot samples are heatin " collect another 4 test tubes and label them A1" B1" 61 7 D1# Place an identical ,olume of distilled water in each of the test tubes A1" B1" 61 7 D1# $he ,olume of water must be enou h to co,er the beetroot samples" but this time it is important that the ,olume of water is the same in each tube# $ransfer the beetroot c!linders to their correspondin tubes A1" B1" 61 7 D1# $he c!linders will now be co,ered b! distilled water# 5ea,e these samples in a test tube trac' for a fi*ed period of 14 minutes at room temperature# &Durin this time the pi ment betac!anin should diffuse throu h the dama ed membranes of the beetroot cells# $he more denatured are the proteins" the faster will be the rate of diffusion of the betac!anin#( After 14 minutes carefull! ta'e out the beetroot c!linders and throw them awa!# <eep the water= $he water contains the pi ment which will contain an! pi ment that has lea'ed out of the tissue" throu h the denatured proteins and is what !ou will assess#

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Year 10 2013-2014 Pre-Diploma Biolo !

OBTAINING RESULTS AND TREATING THEM $he intensit! of the colour of the solutions shows how much pi ment is present# >a'e a scale from 1 to 4 which could be used to estimate the intensit! of the red pi ment# $his is a %uantitati,e scale and is better than simpl! describin the colour# Write down the scale you decide to use# ?ow use the scale to estimate the intensit! of the pi ment in the water in each test tube# $ake a table of your results. You can confirm the accurac! of !our scale with ,er! accurate readin s from a colorimeter" usin a blue filter &complementar! to red(# /f there is time" use the colorimeter to confirm !our scale of results# %lot the results you obtain on a graph. >a'e a line raph @ it will not ha,e man! plots" but the raph should pro,ide a cur,e from which !ou can see the appro*imate temperature at which the escape of betac!anin suddenl! accelerated# At the temperature where the proteins are denatured and can no lon er control diffusion" there will be a reater densit! of the pi ment# CONCLUSIONS omment upon the results and the graph which you obtained# +hat does the raph show? +hat happened? +hat is the e*planation? /f the temperature ,alue for the denaturin of the membrane proteins is different from the one proposed in !our h!pothesis" comment on the possible e*planations b! anal!sin !our e*perimental method" or re,iewin the h!pothesis#

Technician: each group probably requires: Beetroot Cork borer 8 boiling (test) tubes Equipment for water baths, incl. ice boiling water $istille" water

Scalpel Scissors Forceps !ra"uate", measuring c#lin"er Cutting surface

Aohn )sborne April 2014

Year 10 2013-2014 Pre-Diploma Biolo !