Organization of Biochemistry I and II labs

The first semester of this course is designed to introduce you to methodologies and techniques used in modern biochemistry. You will learn these methods while studying a protein of interest (POI) that has been selected and assigned to you. Unlike previous lab courses, pre-canned protocols will not be provided. This laboratory is designed for you to learn to explore, research, and investigate science independently. You are expected to read textbooks, literature, and instructions before lectures and lab sessions. The lab and lecture are designed to help you teach yourself—they are not intended to passively feed you information.

The assigned POIs have been studied previously by students in CHEM4411/4421. These proteins, which are all aminotransferase enzymes, have demonstrated activity in vitro. You will study the protein’s structure and active site in order to understand what amino acid (AA) residues in the active site confer specificity for particular substrates. You will then propose and generate a site-specific AA mutation that you predict would alter that specificity. In the second semester (CHEM4421), you will examine the function of your POI and the mutant that you created in the Fall semester. All of the material you study in the Fall semester is relevant to the second semester. This guide and the data you acquire are the information that you will heavily rely on in the second semester. Because each protein is different, the experiments for each POI group will be different. In the second semester, you will design and execute your own experiments in order to characterize how the mutation affected your POI. 

All of the above is genuine (original) research, and in this way may differ from the experiments that you performed in your previous undergraduate lab courses. Any characterization that you accomplish for your POI should be considered as genuine scientific progress (such as in the publications you read). If your data and results are reliable and thorough, they may be published or disseminated via databases (e.g., TOPSAN) with you as an author.

Performing research of this sort is an excellent way to learn how to approach a question scientifically. However, because it is truly original research, results are not guaranteed. Your grade will not rely on you getting a specific set of data! However, regardless of how well your experiments work, you will be responsible for making reasonable and logical choices in your approach, and for making a concerted and sustained effort to move your project along.