Chapter 24, Problem 5EQ

Chapter 21 describes a method known as Western blotting that can be used to detect a polypeptide that is translated from a particular mRNA. In this method, a particular polypeptide or protein is detected by an antibody that specifically recognizes a segment of its amino acid sequence. After the antibody binds to the polypeptide within a gel, a secondary antibody (which is labeled) is used to visualize the polypeptide as a dark band. For example, an antibody that recognizes α-galactosidase A could be used to specifically detect the amount of α-galactosidase A protein on a gel. The enzyme α-galactosidase A is defective in individuals with Fabry disease, which shows an X-linked recessive pattern of inheritance. Amy, Nan, and Pete are siblings, and Pete has Fabry disease. Aileen, Jason, and Jerry are brothers and sister, and Jerry has Fabry disease. Amy, Nan, and Pete are not related to Aileen, Jason, and Jerry. Amy, Nan, and Aileen are concerned that they could be carriers of a defective α-galactosidase A gene. A sample of cells was obtained from each of these six individuals and subjected to Western blotting, using an antibody against α-galactosidase A. Samples were also obtained from two unrelated normal individuals (lanes 7 and 8). The results are shown here.

(Note: Due to X-chromosome inactivation in females, the amount of expression of genes on the single X chromosome in males is equal to the amount of expression from genes on both X chromosomes in females.)

A. Explain the type of mutation (e.g., missense, nonsense, promoter, etc.) that caused Fabry disease in Pete and Jerry.

B. What would you tell Amy, Nan, and Aileen regarding the likelihood that they are carriers of the mutant allele and the probability of having affected offspring?