Solution Set 4_2011 - Bioc100B Winter 2011 Rubin Solution...

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Bioc100B Winter 2011 Rubin Solution Set #4 Due Tuesday, February 8 th 4:00 P.M. 1) Derive the Michaelis-Menten equation for a mixed inhibitor (an inhibitor that binds both E and ES). You can assume that substrate binding to EI is negligible.
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Bioc100B Winter 2011 Rubin
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Bioc100B Winter 2011 Rubin 2) Propose a kinetic assay that could be used to measure the K I of the noncompetitive kinase inhibitor Gleevec. Some considerations: What are the known quantities in your enzyme catalyzed reaction? What is the data you actually measure and how do you measure it? You will need to research techniques for how biochemists can monitor progress of a kinase reaction in the laboratory. How do you plot this data for analysis? Sketch examples. What kinetic constants do you extract from analysis of the data and how do you calculate them? How can you vary the reaction conditions (i.e. known quantities in the reaction) to manipulate the kinetic constants and derive K I?
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Bioc100B Winter 2011 Rubin Note: Another common way to measure phosphorylated product is to use ATP in the reaction that contains a P 32 radiolabel in its gamma phosphate. Then one can detect the transferred phosphate either by scintillation counting (agter purification of the protein from the reaction) or on an SDS-PAGE gel with Phosphorimaging.
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Bioc100B Winter 2011 Rubin
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Bioc100B Winter 2011 Rubin 3) Read the recent review article by James Robertson entitled “Mechanistic Basis of Enzyme- Targeted Drugs” in the journal Biochemistry. The reference is as follows: Robertson, J.G. Biochemistry, Volume 44, Issue 15, pages 5561-5571, 2005. You can access the journal online and download the paper from a computer with a ucsc.edu address (for example, computers in the Science and Engineering library). a) From his survey Robertson concludes that a majority of enzyme-targeting drugs have structures that resemble substrates. i) Give two examples of such substrate mimics, describe which enzymes they inhibit,
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This note was uploaded on 03/16/2011 for the course BIOC 100B taught by Professor Sethrubin during the Winter '10 term at University of California, Santa Cruz.

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Solution Set 4_2011 - Bioc100B Winter 2011 Rubin Solution...

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