Title: Sickle Cell Anemia
1Sickle Cell Anemia
- An example of why
- a change in protein can lead to disease
- a change in DNA can lead to a change in protein
2Ground Rules for Class Discussions and Workshops
- Be on time.
- Speak so that everyone from front to back can
hear you. - Listen when others are speaking.
- If its review for you, use you intellect to hear
it in a new way. - Write down your answers or consolidate to print.
3Central Dogma
DNA
RNA
Protein
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5What do you already know about hemoglobin?
- What is the function of hemoglobin?
- What class of biomolecules does hemoglobin belong
to? - What are the symptoms of sickle cell anemia?
- Is sickle cell anemia hereditary?
- What does that tell us?
6Proteins
- synthesized from amino acids
7Circle Triangle Square Bond Amino terminal
Carboxy terminal
84 classes of structure.
9Website for Amino acid interactive Workshop
- Amino acids everyone open to this page
- http//iws.ohiolink.edu/chemistry/biochemistry/aat
ut.html
10Power of the R Groups
- Note the one letter and 3 letter abbreviations
for you amino acid(s). - Identify the atoms in red, blue, white, gray, and
other colors - Find the carboxy group, amino group, beta carbon,
R group - Categorize the amino acids and be able to say
why some fit in more than one category! - Aromatic
- Aliphatic, unbranched
- Aliphatic, branched
- Polar
- Positively charged (basic)
- Negatively charged (acidic)
- Small
- Has a sulfur atom in the R group
- Hmm?
- Which are hydrophobic vs. hydrophilic?
- Which would attract each other if brought
together? - Which would repel?
- Which would likely fold to the interior in an
aqueous environment? - Which would likely fold to the exterior in a
lipid environment?
11The Nucleic Acids DNA and RNA
- DNA
- synthesized from deoxynucleotide triphosphates
(dNTPs) - RNA
- synthesized from nucleotide triphosphates
- (NTPs)
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13OH
5 and 3
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15Website for interactive workshop for DNA analysis
- DNA sequence
- Write the primary sequence of the DNA displayed
in 3B from the 5 to the 3 end of both strands
16Central Dogma
DNA
Transcription
RNA
Translation
Protein
17DNA
RNA (with ribosomes)
18Translation
19Translation exercise
- Translate the following sequence using the codon
table - ATG GTG CAC CTG ACT CCT GAG GAG AAG TCT GCC GTT
ACT - Perform same procedure on the sequence below
using a software program - ATG GTG CAC CTG ACT CCT GTG GAG AAG TCT GCC GTT
ACT http//us.expasy.org/tools/dna.html - How many nucleotides have changed in the codon in
boldface? - What is the amino acid difference in the two
sequences? - What is the quality of that difference with
respect to R groups?
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21 22The early evidence that sickle cell anemia is
caused by an amino acid change in hemoglobin.
Tryptic digest the protease trypsin cleaves C
terminal to lysine and arginine.
23Summary
DNA (mutated changed)
RNA (mutated)
Protein (mutated)
Remember Mutation is not always bad! For
example Mutation ? Evolution ? An
additional normal genome
24Multiple sequence alignment for cytochrome C
mutation and conservation
- Human protein accession number AAA35732 (see next
slide) - Dog protein accession number XP_532493
- Yeast protein number from structure database
- 1YCC
- CLUSTAL W PROGRAM
25FASTA format for a protein sequence in single
letter code
Hemoglobin HBB1 gtgi4504349refNP_000509.1
beta globin Homo sapiens MVHLTPEEKSAVTALWGKVNVDE
VGGEALGRLLVVYPWTQRFFESFGDLSTPDAVMGNPKVKAHGKKVLGAFS
DGLAHLDNLKGTFATLSELHCDKLHVDPENFRLLGNVLVCVLAHHFGKEF
TPPVQAAYQKVVAGVAN ALAHKYH
26FASTA format for a protein sequence in single
letter code
gtgi1244762gbAAA98563.1 p53 tumor suppressor
homolog MSQGTSPNSQETFNLLWDSLEQVTANEYTQIHERGVGYEYHE
AEPDQTSLEISAYRIAQPDPYGRSESYD LLNPIINQIPAPMPIADTQNN
PLVNHCPYEDMPVSSTPYSPHDHVQSPQPSVPSNIKYPGEYVFEMSFAQ
PSKETKSTTWTYSEKLDKLYVRMATTCPVRFKTARPPPSGCQIRAMPIYM
KPEHVQEVVKRCPNHATAKE HNEKHPAPLHIVRCEHKLAKYHEDKYSGR
QSVLIPHEMPQAGSEWVVNLYQFMCLGSCVGGPNRRPIQLV FTLEKDNQ
VLGRRAVEVRICACPGRDRKADEKASLVSKPPSPKKNGFPQRSLVLTNDI
TKITPKKRKIDD ECFTLKVRGRENYEILCKLRDIMELAARIPEAERLLY
KQERQAPIGRLTSLPSSSSNGSQDGSRSSTAFS TSDSSQVNSSQNNTQM
VNGQVPHEEETPVTKCEPTENTIAQWLTKLGLQAYIDNFQQKGLHNMFQL
DEFT LEDLQSMRIGTGHRNKIWKSLLDYRRLLSSGTESQALQHAASNAS
TLSVGSQNSYCPGFYEVTRYTYKHT ISYL
27How to prepare the sequences for the MSA on
ClustalW
- For Human and Dog
- Go to NCBI
- Select to search the protein database from the
dropdown menu - Enter the Accession Number (previous slide) and
GO - Click on the link
- Change the display to a FASTA file
- Copy the FASTA output for both species into a
single text file. Make sure the header is
separate from the sequence. - For Yeast
- Clink on the link, find the FASTA format and copy
into the same file - Copy or upload the file into ClustalW
28Workshop due as hardcopy when you arrive Thursday
AM.
- Include answers from within todays class. Email
to me by 9 AM Wed. - Print out your ClustalW results and attach a
short paragraph discussing how Clustal W gives
you a clue as to which part(s) of the Cytochrome
C protein you would hypothesize are most
important to its function (which is/are the same
in all 3 organisms). Start your paragraph as a
hypothesis as to which parts are most important,
and write your discussion as a defense of your
hypothesis. - Find out the chromosomal location of the gene
that causes sickle cell anemia. - Give the name of the gene.
- State the nucleotide change and amino acid change
that leads to sickle cell anemia (there may be
more than one change that gives rise to the
disease) - If sickle cell anemia is so devastating, why has
it lasted in the population for such a long time?
Give a molecular, mechanistic, evolutionary
explanation (you may have to do a little research
to get this). What does the sickled molecule do
that the normal molecule cant?