17 Aug Question: A Female Patient (age, 56) Has Been Diagnosed With Breast Cancer. A Sample Of The Tumor As Well As Surrounding Normal Tissue Have Been Biopsied And Grown In Cell Culture For Further Analysis. Molecular Profiling Analysis Using A DNA Microarray Chip Reveals That The Gene For Protein Kinase X (PKX) Is Highly Overexpressed. Your Hypothesis Is That This …
A female patient (age, 56) has been diagnosed with breast cancer. A sample of the tumor as well as surrounding normal tissue have been biopsied and grown in cell culture for further analysis. Molecular profiling analysis using a DNA microarray chip reveals that the gene for Protein Kinase X (PKX) is highly overexpressed. Your hypothesis is that this may be causing the uncontrolled cell growth in her tumor. PKX is known to phosphorylate a second protein called c-SRC (“sarc”) and activate it. Remembering your cell biology training from a decade ago, you realize that you cannot draw the conclusion that PKX is involved in her tumor growth without characterizing the enzyme kinetics for PKX.
You make a cell extract from the tumor cells and a separate extract from her normal cells grown in culture. Both extracts are derived from the same number of cells. A constant amount of tumor cell extract is added to a series of tubes containing increasing quantities of c-SRC. You allow the reaction to proceed only long enough to be able to measure the amount of c-SRC that has been phosphorylated and then terminate the reaction so that initial rate conditions are satisfied. You repeat the reaction for the normal cell extract using a different series of tubes containing the same increasing quantities of c-SRC.
Assume for a moment that the enzyme from the tumor cells was found to contain a mutation in its active site. This in turn caused the tumor enzyme to have a much higher kcat value than the normal enzyme. As a result, the kcat cannot be discounted from any kinetic calculation. How would this influence the enzyme kinetics?
1.The Vmax and Km of the tumor enzyme would both be higher than those for the normal enzyme.
2.The Vmax and Km of the tumor enzyme would both be lower than those for the normal enzyme.
3.Only the Km would be lower for the tumor enzyme than that for the normal enzyme.
4.Only the Vmax would be higher for the tumor enzyme than that for the normal enzyme.
5.The tumor enzyme would exhibit sigmoidal kinetics compared to a square hyperbola for the normal enzyme.
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