Chapter 5

Chapter 5 - Chapter 5: Protein...

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Chapter 5: Protein Purification/Characterization Protein Purification o Getting proteins out of the cell Bacteria harder to open tough cell wall High pressure Sonication Grind with sand or alumina Mammalarian cells Homogenize High pressure/pressure change o When breaking cells open pH ionic strength osmotic pressure temperature Inhibit proteases redox environment vessels-Do proteins stick? some need cofactors others don’t Proteases-proteins that chew up other proteins Differential centrifugation Separate cell compartments Select compartment where enzyme is located o Membranes dissolved in some type of detergent difficult to deal with Following your protein during purification Enzymatic assay: protein converts A B Linear region-measure of your enzyme present
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Unit of activity: defined amount of product produced under a specific set of conditions based on assay. Eg.
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This note was uploaded on 09/03/2008 for the course BIOL 430 taught by Professor Lspremulli during the Fall '08 term at UNC.

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Chapter 5 - Chapter 5: Protein...

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