Name Class Date

Amino Acid Sequences: Indicators of Evolution

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Problem
How can you use proteins to determine how closely organisms are related?

Introduction
Biologists have many ways to study evolution. They can use fossils to learn about ancient species. They can compare
the anatomy of modern species. They can observe the order in which cells develop in embryos. All these clues reflect
what took place over time at the molecular level. DNA and proteins, the genes and the products of genes, provide
powerful evidence for descent with modification.

As DNA changes over time, the proteins that are produced by the DNA change too. The result is that many organisms
have similar, but not identical, versions of a given protein. Differences among these homologous proteins provide clues
to evolution.

In Part A of this lab, you will compare amino acid sequences of hemoglobin from eight mammals. In Part B, you will
analyze data about sequences in a second protein—cytochrome c. In Part B, the organisms will be more diverse.
Skills Focus Materials
Analyze Data, Graph, Draw Conclusions • highlighter pen, light-colored
• graph paper

Pre-Lab Questions
1. Predict Based only on their anatomy, rank gorillas, bears, chimpanzees, and mice from most recent common ancestor
with humans to least recent common ancestor.
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2. Use Analogies You tell a story to a second person who tells it to a third person, and so on. As the story is retold,
changes are introduced. Over time, the number of changes increases. How is this process an analogy for what happens
to DNA over time?
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3. Infer Hemoglobin from two species is compared. On the long protein chains, there are three locations where the
amino acids are different. Where would you place the common ancestor of the two species on the “tree of life,” and
why?
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Procedure
Part A: Comparing Amino Acid Sequences in Hemoglobin
Hemoglobin is the molecule in blood that carries oxygen. This complex molecule contains four protein chains. Figure 1
shows the amino acid sequence for one of those chains in eight mammals. Each letter stands for a different amino acid.
Each column is a location on the protein chain. NOTE: Locations where the amino acids are identical in all eight
mammals are not shown.

1. Use the row labeled Human as your control. Compare the sequence for the bear to the sequence for humans. When
you find a difference in the bear sequence, highlight it.
2. Repeat Step 1 for each of the other mammals. Be sure to compare each sequence to the sequence for humans.

4 5 6 9 10 12 13 20 25 33 41 43 50 51 52
Human T P E S A T A V G V F E T P D
Bear T G E S L T G V G V F D S A D
Chimpanze
T P E S A T A V G V F E T P D
e
Gibbon T P E S A T A V G V F E T P D
Gorilla T P E S A T A V G V F E T P D
Monkey T P E N A T T V G L F E S P D
Mouse T D A A A S C S G V Y D S A S
Shrew S G E A C T G E A V F D S A S

54 56 58 68 69 70 71 72 73 75 76 77 80 87 104
Human V G P L G A F S D L A H N T R
Bear I N P L N S F S D L K N N K K
Chimpanzee V G P L G A F S D L A H N T R
Gibbon V G P L G A F S D L A H N Q R
Gorilla V G P L G A F S D L A H N T K
Monkey V G P L G A F S D L N H N Q K
Mouse I G A I T A F N D L N H S S R
Shrew V G P L H S L G E V A N N K R

11
109 110 112 116 117 118 121 125 126 130 139
5
Human V L C A H H F E P V Y N
Bear V L C A H H F E Q V Y N
Chimpanze
V L C A H H F E P V Y N
e
Gibbon V L C A H H F E Q V Y N
Gorilla V L C A H H F E P V Y N
Monkey V L C A H H F E Q V Y N
Mouse M I I G H H L D A A F T
Shrew V L V A S K F E P V F N

2
Figure 1 Comparison of amino acid sequences in hemoglobin from eight mammals

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3. In the space below, make a data table to record the number of differences you found for each mammal in
comparison to humans.

Part B: Differences in Cytochrome c

Cytochrome c takes part in electron transport during the last stage of cellular respiration. This enzyme can be found
in bacteria, yeasts, fungi, plants, and animals. The human cytochrome c molecule is relatively small. Its single strand
of protein has 104 amino acids.
4. In Figure 2, human cytochrome c is the standard. Column 1 lists the species that are being compared to humans.
Column 2 lists the number of differences for each pairing. Use the data to make a bar graph on a separate sheet of
graph paper. Select an order for the bars that will best reveal a pattern in the number of differences.
Figure 2 How human cytochrome c differs from cytochrome c found in other species

Species Number of Differences

Chimpanzee 0
Fruit fly 29
Horse 12
Pigeon 12
Rattlesnake 14
Red bread mold 48
Rhesus monkey 1
Snapping turtle 15
Tuna 21
Wheat 43

4
Analyze and Conclude
1. Draw Conclusions Based on the hemoglobin data, which mammal listed is most closely related to humans? What is
the evidence for your conclusion?
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2. Analyze Data Does the cytochrome c data support your conclusion in Question 1? Explain.
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3. Evaluate Does the data support the rankings you made as part of the pre-lab? If not, how would you explain any
differences?
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4. Design an Experiment The cytochrome c in both horses and pigeons differs from the human protein at 12 locations.
Based on this data, you might infer that horses and pigeons are closely related. What could you do to support or refute
this hypothesis?
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5. Communicate In terms of descent, what does it mean to say that humans are more closely related to gorillas than to
monkeys?
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6. Interpret Data A student used the hemoglobin data to conclude that mice and shrews are more closely related
than are mice and humans or shrews and humans. Was the student correct?
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