You are studying the binding of proteins to the cytoplasmic face of cultured neu

Question

You are studying the binding of proteins to the cytoplasmic face of cultured neuroblastoma cells and have found a method that gives a good yield of inside-out vesicles from the plasma membrane.  Unfortunately, your preparations are contaminated with variable amounts on right-side-out vesicles.  Nothing you have tried avoids this problem.  A friend suggests that you pass your vesicles over an affinity column made of lectin (binds carbohydrates) couple to beads.  What is the point of your friend's suggestion?

Explanation / Answer

      When the preparations arecontaminated with variable amounts on righr-side-out vesicles andthe suggestion of the       friend to pass thevesicles over an affinity column made of lectin couple to beads isa good suggestion.       This is because, the rightside out vesicles can be removed by this technique.       Initially, to the vesiclesthat are on right side out cetyltrimethyl ammonium tosylate andsodiumdodecyl sulphonate are       added to obtainthermodynamically stable vesicles.       To this thermodynamicallystable vesicles, carbohydrate moieties are linked to thehydrocarbon chain through an N-       glycosyl linkage.       The carbohydrate groupsthat are displayed on the outer surface of the vesicle will bind tothe lectins in the solution       because the lectin acts asa ligand.       Once these vesicles bindto the lectins through carbohydrate, they remain in the column andthe desired protein solution       is eluted out.       So, the point ofsuggestion is that the right side out vesicles will bind to thelectins and the contamination can be       removed.       Hope you are satisfiedwith the answer.

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