Question: In class, you have analyzed the experiments that have shown that the protein α. (wild type) is my…


Myristoylation and palmitoylation
In class, you have analyzed the experiments that have shown that the protein α. (wild type) is myristoylated and palmitoylated. The next question that the researchers wanted to address was whether the palmitoylation of the protein was required for its membrane association The experiment was done as follows: Plasmids carrying the gene corresponding to ao and to the mutant a, C3A were transfected into COS-7 cells. After two days, cells were radiolabeled with [hS)methionine Then, the cells were harvested and centrifuged at 100,000xg. Two fractions were obtained: supernatant (S2) and pellet (P2). The αο and the mutant α.c3A proteins that had been labeled with Ps5]methionine were immunoprecipitated from both fractions. The presence of wild type and mutant a, was analyzed by electrophoresis and exposure of the gel to a film. The results are shown in the figure below: P2 S2 P2 S2 Compare lanes 1 and 2 with lanes 3 and 4 1. Does the mutation affect the subcellular localization? Is palmitoylation of Gag required for membrane association? Explain your answer.
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In class, you have analyzed the experiments that have shown that the protein α. (wild type) is myristoylated and palmitoylated. The next question that the researchers wanted to address was whether the palmitoylation of the protein was required for its membrane association The experiment was done as follows: Plasmids carrying the gene corresponding to ao and to the mutant a, C3A were transfected into COS-7 cells. After two days, cells were radiolabeled with [hS)methionine Then, the cells were harvested and centrifuged at 100,000xg. Two fractions were obtained: supernatant (S2) and pellet (P2). The αο and the mutant α.c3A proteins that had been labeled with Ps5]methionine were immunoprecipitated from both fractions. The presence of wild type and mutant a, was analyzed by electrophoresis and exposure of the gel to a film. The results are shown in the figure below: P2 S2 P2 S2 Compare lanes 1 and 2 with lanes 3 and 4 1. Does the mutation affect the subcellular localization? Is palmitoylation of Gag required for membrane association? Explain your answer.

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