2. You are interested in receptor protein tyrosine kinases and you are purifying

Question

2. You are interested in receptor protein tyrosine kinases and you are purifying a novel phosphotyrosine-binding to characterize this protein you need to protein (NPBP-1). In order in your partially purified separate it from three other proteins p,Y,and material. The proteins in your preparation have the following properties: Size (kDa) pl binds to phosphotyrosine Freehonatan protein X protein Y Yes 29 129 Z no 1 protein NPBP-1 For each answer below, you must explain why the technique separates the NPBP-1 from the other protein (theory), which protein is retained on the column longer, how the retained proteins are eluted from the column, etc. you are planning on using an ion exchange column, you must specifically state which type (cation or anion) and pH conditions. What type of separation technique can be used to separate protein NPBP-1 from protein z? e would -then have XY- NPB 1 What type of separation tec can be used to separate protejp1NPBP-1 from protein x? NPB what type of separation technique can be used to separate protein NPBP-1 from protein Y? polar Probably ubine a TLC plate f

Explanation / Answer

The methods suggested in the answers don't work.
To separate NPBP-1 from protein X Size Exclusion chromatography which is used to separate large molecules
of proteins.
To separate NPBP-1 from protein Y , Ion Exchange chromatography is ideal which separates ions in proteins and polar molecules based on their affinity to the ion exchanger that may be cationic or anionic exchanger.

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