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16 Objectives for Weeks 5-9
- Develop kinetic assay and determine catalytic constants for wild-type and ncAA-mutant enzymes (see below for advice on running assays).
- Modify assay as necessary to obtain meaningful, reproducible results. Purify additional protein if necessary.
- Obtain kinetic parameters for wild-type and ncAA-mutant enzymes so that comparisons can be made, both to literature values as well as among the enzymes tested in this experiment.
- Design and execute individual assays for protein structure and function.
- Based on the known structure of the enzyme, rationalize the obtained results and attempt to determine the structure-function relationship of that particular site.
- Design and run experiments to test your hypotheses on structure and function
- Mass spec analysis of purified proteins to confirm expected composition-if mass spec time is available to lab class.