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AP Biology
Mrs. Schulte
Experiment #1
The effect of various chemicals on proteins and their form and function
Abstract:
In this experiment various ways to denature a protein were tested. In part one various
strong acids and bases were added to a protein solution and caused a overall reaction that
eventually lead to coagulation the more that was added. The addition of strong acids or bases
affects the number of charges on amino acid side chains and interferes with ionic salt bridge
formation in proteins. Strong acids increase the concentration of H+ ions in solution which
neutralize negatively charged side chains. Strong bases increase the concentration of OH- ions
that in turn neutralize positively charged side chains. Proteins have an optimal pH range where
they are most stable, soluble, and active small changes in this range can cause reversible changes
to the protein; as seen in the denaturation of albumin by ammonium sulfate.
We also tested the effect of various inorganic and organic additive to a protein solution
and found some extreme changes in color and various amounts of precipitate. Polar organic salts
such as alcohols (isopropyl alcohol) and acetone interfere with hydrogen bonding and cause the
protein to lose its shape and essentially its function. The poisonous nature of heavy metal salts
containing Ag+, Hg2+>, and Pb2+ ions such as CuSO4 and AgNO3, is due to protein denaturation
as well.
In addition to that we conducted an experiment to “salt out” a protein with ammonium
sulfate. We were able to take the coagulated protein out of the solution and dissolve it back in
with minimum effect on the protein’s shape. The high concentration of inorganic salts such as,
ammonium sulfate, are used to precipitate proteins without loss of protein activity. The solubility
of a protein decreases as the concentration of the ionic compounds increases and the
concentration of the protein precipitates completely. Salting out results from changes in
hydrogen bonding between water molecules and the protein. Because it involves mild conditions
the process is usually reversible, as seen in our experiment, and can be used to isolate proteins
and purify them to remove contaminants.
Finally we tested the effect changes in temperature have on protein solutions and
concluded that certain temperatures denature the protein due to the excess energy. It destabilizes
all of the major forces that hold a protein together. Proteins have an optimal temperature range
where they still have protein activity which is usually up to 50 degrees celsius.
Results:
Test tube 1 2 3
Results Blue, fizzy, little bit of Really white, A little cloudy, slight
precipitation bubble/foamy yellow tint
Observations
Test tube 2 (Filtrate + CuSO4) Cloudy light blue color, slightly thicker, no
precipitate
Test tube 3 (Redissolved solid + CuSO4) Very clear, medium blue tint, no precipitate,
slight purple
Analysis/conclusion:
In this experiment we were able to conclude that HCl and NaOH do have an increasing
effect on the denaturation of albumin, casein, and gelatin, inorganic/organic additives such as
CuSO4, AgNO3 and isopropyl alcohol have a negative effect on protein activity, ammonium
sulfate can successfully salt out a protein and maintain its activity, and heat outside of the
proteins temperature range will also cause irreversible denaturation. My hypothesis that these
would have a negative effect on the protein activity came true for all of the experiments
excepting salting out a protein because its protein activity remained more or less unchanged.
There could have been a few sources of error, human error in the measuring or recording
informations wrong could possibly skew our results as well as contaminated equipment. Being
able to investigate this in a real life situation, similar to the previous potato lab, would be really
interesting. Seeing how proteins work in the human body would also be very interesting
Sources:
Flinn Scientific. (n.d.). Retrieved December 08, 2017, from http://www.flinnsci.com/