You are purifying the enzyme Hexokinase (which converts Glc rightarrow Glc-6-PO_

Question

You are purifying the enzyme Hexokinase (which converts Glc rightarrow Glc-6-PO_4) from rat liver cells. The figure shows your results from the various purfication steps. Both the SDS-PAGE results and the corresponding Western blot (using a Hexokinase-specific antibody) is shown. Galactose is a competitive inhibitor of Hexokinase. Answer the questions that follow based on these results Can we conclude that we have purified Hexokinase? ? Yes: SDS-PAGE shows only one band after the Galactose affinity column. Yes: The Western blot shows only one band after the Galactose column. Yes: Both the SDS-PAGE and Western blot show one band after the Galactose column. Yes: We know that Galactose binds Hexokinase specifically. We can conclude that most likely, the sample in the Galactose affinity column lane will have the highest specfic activity. This conclusion is based on what data shown? The single band seen in the Western for that condition. The single band seen in the SDS-PAGE for that condition. The thickness of the band in the Western for that condition. The thickness of the bands in the SDS-PAGE. Which lane is most likely to show the lowest activation energy for the Glc rightarrow Glc-6-PO_4 reaction catalyzed by Hexokinase? The total cell lysate. It will have the most amount of total enzyme. The Galactose affinity column purified enzyme. It will have the purest enzyme. All EXCEPT the Galactose affinity purified enzyme will have the same activation energy. Galactose inhibits the reaction. All will have the same activation energy. Enzyme purity does not affect the activation energy of the reaction. I measure the K_m and V_max for Hexokinase in the Total cell lysate and the Galactose column purified sample, using the same volume of each sample. What will be TRUE about the measured K_m and V_max? K_m and V_max will be the same for both. K_m and V_max will be higher in the Galactose column purified sample. K_m will be higher in the Galactose column purified sample. V_max will be the same in both. K_m will be the same in both. V_max will be higher in the Galactose column purified sample.

Explanation / Answer

Ans-1-a- in this part all the four parts are true.

Ans-1-b- in this part a,b and c are true and d is false.

Ans-1-c- in this part c is true.

Ans-1-d- in this part b is true.

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