Question: 5. Compare and Contrast the two strategies below for identifying and characterizing protein- prot…

Reaction Properties that inform utility for a
particular application

      Strategy 1: Protein A
‘ Pull down’

      
      Strategy 2: Yeast
Two-Hybrid

Relative affinity of protein-protein interactions
necessary for successful isolation using this
procedure

Need for cDNA encoding the ‘bait’ protein of
interest

Ability to identify multi-protein complexes

Ability to examine impact of extracellular ligands on
extent and/or affinity of protein-protein interactions

Amenable to ala-scanning mutagenesis to refine
understanding of protein-protein interaction domains