You are tasked with purifying two integral membrane proteins from HEK293 cells: C4: KDELR1, KDEL (Lys-Asp-Glu-Leu) endoplasmic reticulum protein retention receptor 1(-29 kDa) D11: Corf105, androgen-dependent TFPI-regulating protein (-27 kDa) Each expressed protein has a C-terminal 6x-His tag. Use this information to answer the following questions: -a. You must decide between n-Dodecyl B-D-maltoside (DDM, CMC: 0.15 mm. Aggregation number: 98) and N-Octyl-B-D-glucoside (BOG, CMC: 23.4 mm, Aggregation number: 27) as detergents to solubilize the proteins. What factors would you consider in choosing a detergent to solubilize the proteins? b. When you run a initial nickel column to purity C4 and D11 with both detergents, you get the following results: B-OG DDM D11 04 D11 C4 25KDa - B-OG DDM D11 C4 D11 cm - 25KDa Why do both detergents yield similar results? c. You decide to move forward with DMM. The results of your nickel column purification polishing step are as follows: C4 D11 E1 E2 N E3 W2 E3 E2 W2 E1 M LL Why do both detergents yield similar results? You decide to move forward with DMM. The results of your nickel column purification polishing step are as follows: D11 C4 MW2 E1 E2 E3 W2 E1 E2 E3 25KDa Why is imidazole used to elute your proteins from the column? d. As a confirmatory experiment, you decide to assess the ability of spin columns with different molecular weight cutoffs to retain DDM micelles with your protein, you get the following results: Cut off 5k 30k 50k 100k DDM after, ug 28.28737997 24.78986966 22.24093373 0.430064065 Based on the above results, what can you conclude about the micelle molecular weight of DDM? e. Further experiments with DMM are done to assess the change in absorbance associated with increasing concentrations of DMM: Standard Curve, DDM vs Abs 08 07 v115.00032 099456 06 03 04 Abs, on Series Une Series 02 0.1 0 -0.1 DOM Why do you observe the indicated pattern in the data? What is the biophysical phenomenon that explains the data