You are interested in receptor protein-tyrosine kinases and you are purifying a

Question

You are interested in receptor protein-tyrosine kinases and you are purifying a novel phosphotyrosine-binding protein [NPBP-1]. In order to characterize this protein you need to separate it from three other proteins (X, Y, and Z) that are still present in your partially purified material. The proteins in your preparation have the following properties: What type of separtion technique can be used tyo separate protein NPBP-1 from protein X What type of separation technique can be used to separate protein NPBP-1 from protein Y What type of separation technique can be used to separate protein NPBP-1 from protein Z

Explanation / Answer

(a). Technique used to separate protein NPBP-1 from protein X -

- Gel (or size) exclusion chromotography. Since this chromatographic method is used to separate molecules by on the basis of their size. Here the size of proteiz X is much larger than the NPBP-1.

(b). Technique used to separate protein NPBP-1 from protein Y -

- Ion exchange chromotaography. As this chromatography process separates ions and polar molecules based on their affinity to the ion exchanger. Since there is a resonable difference in the isoelectric points of protein Y and NPBP-1, so we will use ion exchange chromatography to separate them.

(c). Technique used to separate protein NPBP-1 from protein Z -

- Affinity chromatography. Since the protein Z does not binds with phosphotyrosine while NPBH-1 binds with it, So we will use Affinity chromatography to separate them.

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