Name _____________________________ Period ___________________ Date __________________
ADDITIONAL INVESTIGATION
Biochemical Evidence for Evolution
Teacher Notes
TIME 45 minutes
TEACHER PREPARATION
STUDENT DIFFICULTY
Purpose Understand how the evolutionary relationships among various
vertebrates are reflected in the similarities and differences between their amino
acid sequences.
Overview Students will analyze amino acid sequences and draw conclusions about
relationships between organisms.
LAB MANAGEMENT
 Review
with students how proteins are formed in the body.
 Complete the first sequence comparison as a group to make sure students
understand how to complete Step 2.
 Table 2 is included as a helpful reference for students so that the abbreviations
used in Table 1 have some meaning.
Post-Lab Discussion Discuss how analyzing DNA and amino acid sequences
allows scientists to learn more about the evolutionary relationships among
organisms. Ask students to consider how this method might be used to determine
relationships among living and extinct species. Explain how sequence comparison
is also used in biomedical and field research by analyzing and comparing protein
sequences with protein databases. Give examples of the value of using DNA and
protein sequences in biomedicine, like the development of new antibiotics to
counteract bacterial resistance to older antibiotics, and cancer research and
inheritance.
OBSERVATIONS
TABLE 3. Matching and Nonmatching Amino Acids
Organism: Chimpanzee; Number of matching amino acids: 96; Number of
nonmatching amino acids: 0. Organism: Fish; Number of matching amino acids:
58; Number of nonmatching amino acids: 38. Organism: Mouse; Number of
matching amino acids: 80; Number of nonmatching amino acids: 16.
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Additional Investigation
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Principles of Evolution
Section 5: Evolutionary Biology Today
Name _____________________________ Period ___________________ Date __________________
Biochemical Evidence for Evolution continued
TABLE 4. Percent Similarity and Difference in Amino Acid Sequence Compared
to Humans
Organism: Chimpanzee; Percent similarity: 100.0; Percent difference: 0.0.
Organism: Fish; Percent similarity: 60.4; Percent difference: 39.6. Organism:
Mouse; Percent similarity: 83.3; Percent difference: 16.7.
ANALYZE AND CONCLUDE
1. Bar graph with "Organism" on the x-axis and "Percent Similarity" on the yaxis; Fish (60.4%), Mouse (83.3%), and Chimpanzee (100%).
2. Chimpanzees are most closely related to humans. Fish are least closely related
to humans.
3. Matching amino acids: 54; nonmatching: 42. Percent similarity: 56.2%,
percent difference: 43.7%.
4. Alanine, Histidine, Alanine, Glycine, Glutamic Acid, Tyrosine, Glycine,
Alanine, Glutamic Acid.
5. Similarity: 14.3%; difference: 85.6%.
6. The gorilla is most closely related to humans. The lemur is least closely related
to humans.
7. They would be very similar.
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Holt McDougal Biology
Additional Investigation
2
Principles of Evolution
Section 5: Evolutionary Biology Today
Name _____________________________ Period ___________________ Date __________________
ADDITIONAL INVESTIGATION
Biochemical Evidence for Evolution
One method scientists use to help determine the evolutionary relationships
between organisms is to analyze and compare the molecular structure of proteins.
Recall that proteins are made up of chains of amino acids. There are 20 amino
acids that are used to make proteins. The particular type and order of the amino
acids in a protein are determined by the DNA code.
To make a given protein, this information is first transcribed into a messenger
RNA (mRNA) code, which is "read," or translated into protein, three nucleotides
at a time. With a few exceptions, each triplet, called a codon, codes for a certain
amino acid. For example, the mRNA codon UCA (that is, uracil, cytosine, and
adenine) codes for the amino acid serine. The code is also somewhat flexible. The
amino acid glycine will be brought into the sequence of a protein if any of the
following codons appear: GGU, GGC, GGA, or GGG.
To better understand ancestral relationships between organisms, scientists
compare the amino acid sequences of the same proteins in different organisms.
Scientists have concluded that the more a sequence from one organism matches
the sequence from another organism, the more closely related the two organisms
are. For example, scientists determined that light-sensing proteins in the brain of
an ancient marine worm are very similar to those found in the vertebrate eye. This
evidence supports the idea that worms and vertebrates share a common ancestor.
Scientists have examined the amino acid sequences of proteins such as
hemoglobin and myoglobin in many organisms and compared them to the
corresponding sequences in humans. Hemoglobin is the protein in red blood cells
that binds to oxygen and carbon dioxide. Myoglobin is a protein found in muscle
tissue. Scientists have used the information from these analyses to strengthen
conclusions drawn from other evidence of the ancestral relationships between
certain organisms.
In this lab, you will compare the amino acid sequences of hemoglobin from
several vertebrate organisms with the amino acid sequence for human
hemoglobin.
PROBLEM
How do the amino acid sequences of vertebrate organisms compare?
MATERIALS
pencil
PROCESS SKILLS
 Analyzing
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Additional Investigation
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Principles of Evolution
Section 5: Evolutionary Biology Today
Name _____________________________ Period ___________________ Date __________________
Biochemical Evidence for Evolution continued
 Calculating
 Inferring
PROCEDURE
1. Table 1 shows part of the amino acid sequence for hemoglobin in several
different vertebrates, including humans. A single-letter abbreviation has been
used to represent each amino acid in the sequences. Table 2 shows which
amino acid each abbreviation stands for.
2. Look at the amino acid sequence of hemoglobin for each organism listed in
Table 1 and compare it to the sequence of the amino acids in human
hemoglobin. Count the number of amino acids that match the human amino
acids in the same position. Record this number in Table 3.
3. Count the number of amino acids that do not match the human amino acids in
the same position. Record the number in Table 3.
4. Calculate the percent similarity between the amino acid sequence of humans
and each of the other organisms using the formula below. Record your results
in Table 4.
Percent Similarity 
number of matching amino acids
 100
total number of amino acids
5. Calculate the percent difference between the amino acid sequence of humans
and each of the other organisms by subtracting the percent similarity from
100%. Record your results in Table 2.
Percent Difference = 100% — Percent Similarity
OBSERVATIONS
TABLE 3. MATCHING AND NONMATCHING AMINO ACIDS
Organism
Number of Matching Amino
Acids
Number of Nonmatching
Amino Acids
Chimpanzee
Fish
Mouse
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Holt McDougal Biology
Additional Investigation
4
Principles of Evolution
Section 5: Evolutionary Biology Today
Name _____________________________ Period ___________________ Date __________________
Biochemical Evidence for Evolution continued
TABLE 4. PERCENT SIMILARITY AND DIFFERENCE IN AMINO ACID SEQUENCES
COMPARED WITH HUMAN SEQUENCE
Organism
Percent Similarity
Percent Difference
Chimpanzee
Fish
Mouse
TABLE 1. HEMOGLOBIN AMINO ACID SEQUENCE FOR VARIOUS VERTEBRATES
Chimpanzee
VLSPADKTN VKAAWGKVG AHAGEYGAE ALERMFLSF
PTTKTYFPHF ALSALSDIH AHKLRVDPV NFKLLSHCL
LVTLAAHLP AEFTPAVHA SLDKF
Fish
SLSDKDKAV VKAIWAKIS PKADEIGAE ALARMLTVY
PQTKTYFSHW GLAALSEIH AFKLRVDPA NFKILSHNV
IVVIAMLFP ADFTPEVHV SVDKF
Human
VLSPADKTN VKAAWGKVG AHAGEYGAE ALERMFLSF
PTTKTYFPHF ALSALSDIH AHKLRVDPV NFKLLSHCL
LVTLAAHLP AEFTPAVHA SLDKF
Mouse
VLSGEDKSN IKAAWGKIG GHGAEYGAE ALERMFASF
PTTKTYFPHF ALSTLSDIH AHKLRVDPV NFKFLSHCL
LVTLASHHP GDFTPAMHA SLDKF
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Holt McDougal Biology
Additional Investigation
5
Principles of Evolution
Section 5: Evolutionary Biology Today
Name _____________________________ Period ___________________ Date __________________
Biochemical Evidence for Evolution continued
TABLE 2. ABBREVIATIONS FOR AMINO ACIDS
Abbreviation
Amino Acid
Abbreviation
Amino Acid
A
Alanine
L
Leucine
R
Arginine
K
Lysine
N
Asparagine
M
Methionine
D
Aspartic Acid
F
Phenyalanine
C
Cysteine
P
Proline
E
Glutamic Acid
S
Serine
Q
Glutamine
T
Threonine
G
Glycine
W
Tryptophan
H
Histidine
Y
Tyrosine
I
Isoleucine
V
Valine
ANALYZE AND CONCLUDE
1. Construct Choose whether a line graph or bar graph best represents the
percent similarity data from Table 4. Construct the graph, showing the rank of
the organisms from least to greatest in terms of percent similarity to the amino
acid sequence of human hemoglobin.
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Holt McDougal Biology
Additional Investigation
6
Principles of Evolution
Section 5: Evolutionary Biology Today
Name _____________________________ Period ___________________ Date __________________
Biochemical Evidence for Evolution continued
2. Analyze Based on the amino acid sequences for hemoglobin, which organism
is most closely related to humans? Which organism is the least closely related
to humans?
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3. Calculate Use the information in Table 1 to calculate the percent similarity
and the percent difference between the amino acid sequence in fish and mice.
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4. Sequence Use Table 2 to determine the amino acids in the following sequence
from a strand of human hemoglobin.
AHAGEYGAE
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5. Calculate The total number of amino acids in the hemoglobin beta-chain is
146. When compared to a human, a lamprey (a jawless fish) has 21 matching
amino acids and 125 nonmatching amino acids. Calculate the percent
similarity and percent difference between humans and lampreys.
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6. Assess Myoglobin is a protein found in the muscle tissue of vertebrates. The
following table contains information about the percent similarity and percent
difference between the myoglobin of various organisms and human
myoglobin.
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Holt McDougal Biology
Additional Investigation
7
Principles of Evolution
Section 5: Evolutionary Biology Today
Name _____________________________ Period ___________________ Date __________________
Biochemical Evidence for Evolution continued
Organism
Percent Similarity
Percent Difference
Baboon
96.0
4.0
Lemur
85.6
14.4
Gorilla
99.3
0.7
Squirrel Monkey
88.8
11.2
Which of these organisms is most closely related to humans? Which is least
closely related to humans?
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7. Infer How would you expect the mRNA codons that code for the amino acids
that make up hemoglobin to compare between humans and chimpanzees?
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© Houghton Mifflin Harcourt Publishing Company
Holt McDougal Biology
Additional Investigation
8
Principles of Evolution
Section 5: Evolutionary Biology Today