GENETIC ANALYSIS: AN INTEG. APP. W/MAS
GENETIC ANALYSIS: AN INTEG. APP. W/MAS
2nd Edition
ISBN: 9781323142790
Author: Sanders
Publisher: Pearson Custom Publishing
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Chapter 10, Problem 8P

Wild - type β - globin protein is composed of 146 amino acids. A β - globin gene mutant known as Hb Cranston contains 157 amino acids. Partial mRNA sequences of β A and Hb Cranston ( β Cr ) are shown. The numbers indicate amino acid positions. Identify the mutation that causes β Cr , and describe how the mutation leads to a longer than normal β - globin protein chain.

Chapter 10, Problem 8P, 8. Wildtype βglobin protein is composed of  amino acids. A βglobin gene mutant known as Hb Cranston

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When human hemoglobin undergoes a mutation, the mutant protein usually does not replace all of the normal HbA in the red blood cells or erythrocytes of the individual. The erythro- cytes contain mixtures of varying amounts of both HbA and the mutant protein depending on the mutation and the individual. Hb Yakima is a mutant human Hb with an Asp-(B99)His mutation. The diagram on the right shows that Hb Yakima was separated by DEAE-cellulose chromatography from HbA with a 0 – 0.1 M linear gradient of NaCl buffered to pH 8.3. Why is chromatog- raphy carried out at pH 8.3? If the isoelectric point of HbA is 6.85, what is the change in total charge caused by the mutation?How does the change in charge explain the chromatography elution profile of the Hb Yakima/HbA mixture? 1,5 -Hb-A Hb -Yakima 1.0 0.5- 20 40 60 80 00 Fraction number O.D578 nm
Below is the DNA base sequence for the normal protein for normal hemoglobin and the base sequence for        (abnormal) sickle cell hemoglobin:    Normal GGG CTT CTT TTT  Sickle   GGG CAT CTT TTT  A)Transcribe and translate the normal and sickle cell DNA.          B)Identify this as a point or frameshift mutation.  Explain.
The proximal histidine residues have been replaced by glycine residues by mutation of the cloned genes for both the α and β subunits of hemoglobin.  With the tetrameric mutant hemoglobin (all subunits being mutant, α H F8 G, β H F8 G), it was found that the “proximal” coordination bonds to hemes in the mutant protein could be replaced by having the small molecule imidazole in the buffers.  Oxygen binding curves for the tetrameric mutant hemoglobin were measured.    A. The degree of cooperativity in oxygen binding for the mutant hemoglobin (with imidazole present) would be expected to   1)  increase 2)  decrease 3)  not be affected)   compared with the normal protein.   B. Justify your answer to part A in terms of what you know about the structural basis of cooperativity in hemoglobin.      C. How would the Hill coefficient for the mutant be expected to change compared with nH for normal hemoglobin, which is ~3?
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