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Guided Inquiry
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Chapter 16 Lab Amino Acid Sequences:

Indicators of Evolution
Problem Big Idea
Molecular biology provides
How can you use proteins to determine how closely organisms evidence for evolution.
are related? Skills Objectives
Students will be able to
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Introduction sequences of proteins.
Biologists have many ways to study evolution. They can use fossils t
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to learn about ancient species. They can compare the anatomy of proteins.
modern species. They can observe the order in which cells develop
Class Time
in embryos. All these clues reflect what took place over time at the 
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molecular level. DNA and proteins, the genes and the products of
Group Size
genes, provide powerful evidence for descent with modification. Individuals
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 varied.

Skills Focus
Analyze Data, Graph, Draw Conclusions

Build Vocabulary Build Vocabulary

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Term Definition successful lab experience if
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compare To examine two or more things in order to discover
ways in which they are alike, or comparable, and
ways in which they are different
sequence The order in which things are arranged, actions are
carried out, or events happen
related Connected by a common, or shared, origin;
descended from a common ancestor
identical Exactly the same or alike in every way

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Advance Preparation Materials

Collect and display images
of the organisms listed in • highlighter pen, light-colored
Figure 1.
• 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.
Students are likely to say that chimpanzees and gorillas have a more
recent common ancestor with humans than do bears and mice. Do not
correct the rankings at this point.

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?
Students can compare retelling the story to the replication of DNA and
the changes in the story to mutations. Despite the difference in time
frame, with both the story and the DNA the number of changes will
increase with time.

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?
The common ancestor should be placed relatively recently along the
tree based on the limited differences between the two proteins and the
time required for mutations to occur.

Teaching Tip Procedure

If your students have not done
the Chapter 13 lab, “From Part A: Comparing Amino Acid Sequences in Hemoglobin
DNA to Protein Synthesis,”
they may be curious about
Hemoglobin is the molecule in blood that carries oxygen. This
or confused by the symbols complex molecule contains four protein chains. Figure 1 shows the
in Figure 1. Refer them to the amino acid sequence for one of those chains in eight mammals.
table on page 79, which maps
single-letter symbols to amino
Each letter stands for a different amino acid. Each column is a
acids. location on the protein chain. note: Locations where the amino
acids are identical in all eight mammals are not shown.

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1. Use the row labeled Human as your control. Compare the Step 2
The rows with the data for
sequence for the bear to the sequence for humans. When you humans are shaded to remind
find a difference in the bear sequence, highlight it. students that humans are the
2. Repeat Step 1 for each of the other mammals. Be sure to control for this activity.

compare each sequence to the sequence for humans.

Figure 1 Comparison of amino acid sequences

in hemoglobin from eight mammals

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
Chimpanzee T P E S A T A V G V F E T P D
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

109 110 112 115 116 117 118 121 125 126 130 139
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
Chimpanzee V L C A H H F E P V Y N
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

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3. Use the data table to record the number of differences you

found for each mammal in comparison to humans.
Sample Data
Data Table
Mammal Number of Differences
in Hemoglobin
Bear 15

Chimpanzee 0

Gibbon 2

Gorilla 1

Monkey 8

Mouse 29

Shrew 24

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 yeasts,
bacteria, fungi, plants, and animals. The human cytochrome c
molecule is relatively small. Its single strand of protein has
104 amino acids.
Step 4 4. In Figure 2, human cytochrome c is the standard. Column 1
Labeling the axes will be easier lists the species that are being compared to humans. Column 2
if students use the y-axis for
the species and the x-axis for lists the number of differences for each pairing. Use the data to
the number of differences. make a bar graph on a separate sheet of graph paper. Select an
Teaching Tip order for the bars that will best reveal a pattern in the number
This lab uses limited data to of differences.
demonstrate one method
biologists use to study Figure 2 How human cytochrome c differs from
evolution. Remind students cytochrome c found in other species
that scientists do not generally
base their conclusions on such
limited data. 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

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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?
The chimpanzee is most closely related. There are no differences between the amino
acid sequences in chimpanzees and humans.

2. Analyze Data Does the cytochrome c data support your conclusion in

Question 1? Explain.
Yes, because there are no differences in the amino acid sequence between
chimpanzees and humans.

3. Evaluate Does the data support the rankings you made in Pre-Lab
Question 1? If not, how would you explain any differences?
Analysis of biochemical differences can reveal relationships between species that are
not obvious based only on anatomy.

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 reject this hypothesis?
You would compare the amino acid sequences in the pigeon and the horse to each
other rather than comparing them to the human protein.

5. Communicate In terms of descent, what does it mean to say that humans

are more closely related to gorillas than to monkeys?
It means that the common ancestor of gorillas and humans lived more recently than
the common ancestor of monkeys and humans.

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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?
No. The number of differences between the hemoglobin for mice and shrews (33)
is greater than the number of differences for mice and humans (29) or shrews and
humans (24).

Build Science Skills

More than 100 locations were not listed in Figure 1 because the amino
acid was the same in all eight species. One possible explanation is that no
mutations occurred in the codons for those locations. Suggest another possible
explanation. Hint: Think about why some mutations are harmful.
Sample answer: Mutations did occur, but the protein that was produced could no longer
perform its function or its ability to perform the function was limited. Individuals with
such a mutation would be less likely to survive and reproduce.

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