Unformatted text preview: Page 6 Name 4. You hypothesize that the CF cell has defective chloride efﬂux because a mutation results in a CFTR protein
that is missing a 20,000 d domain that is required for proper CFTR integration into the plasma membrane.
Therefore, a smaller, non-functional version of CFTR accumulates in the cytoplasm of CF cells. To test your hypothesis you begin by analyzing a total protein extract of both cell types by SDS—PAGE. (8 pts) a. Sketch and label the Coomassie Blue banding pattern you would expect to see if your hypothesis is correct. N b. Suppose that the mutant CF cells are producing a defective CFTR protein that accumulates in the cytoplasm
so that CF cells have 10~times the normal concentration of mutated protein compared to CFTR in normal cells. Sketch and label the banding pattern you would expect to see if this is true. N c. To more thoroughly characterize the proteins in the normal and CF cells, in two separate experiments you:
1) subject both cell types to an ionic wash prior to SDS-PAGE analysis, and 2) subject both cell types to a brief trypsin treatment prior to SDS-PAGE analysis. Sketch and label the results you would expect to see with Coomassie Blue staining of the gels. 10M ' ‘.
wash (1)2?!“ afﬁx“ ...
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- Spring '08
- cell biology